Tango 294, a lipase-like protein

ABSTRACT

The invention provides isolated nucleic acids encoding a variety of proteins having diagnostic, preventive, therapeutic, and other uses. These nucleic and proteins are useful for diagnosis, prevention, and therapy of a number of human and other animal disorders. The invention also provides antisense nucleic acid molecules, expression vectors containing the nucleic acid molecules of the invention, host cells into which the expression vectors have been introduced, and non-human transgenic animals in which a nucleic acid molecule of the invention has been introduced or disrupted. The invention still further provides isolated polypeptides, fusion polypeptides, antigenic peptides and antibodies. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided. The nucleic acids and polypeptides of the present invention are useful as modulating agents in regulating a variety of cellular processes.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of co-pending U.S. patent application Ser. No. 09/333,159, filed Jun. 14, 1999.

BACKGROUND OF THE INVENTION

The molecular bases underlying many human and animal physiological states (e.g., diseased and homeostatic states of various tissues) remain unknown. Nonetheless, it is well understood that these states result from interactions among the proteins and nucleic acids present in the cells of the relevant tissues. In the past, the complexity of biological systems overwhelmed the ability of practitioners to understand the molecular interactions giving rise to normal and abnormal physiological states. More recently, though, the techniques of molecular biology, transgenic and null mutant animal production, computational biology, pharmacogenomics, and the like have enabled practitioners to discern the role and importance of individual genes and proteins in particular physiological states.

Knowledge of the sequences and other properties of genes (particularly including the portions of genes encoding proteins) and the proteins encoded thereby enables the practitioner to design and screen agents which will affect, prospectively or retrospectively, the physiological state of an animal tissue in a favorable way. Such knowledge also enables the practitioner, by detecting the levels of gene expression and protein production, to diagnose the current physiological state of a tissue or animal and to predict such physiological states in the future. This knowledge furthermore enables the practitioner to identify and design molecules which bind with the polynucleotides and proteins, in vitro, in vivo, or both.

The present invention provides sequence information for polynucleotides derived from human and murine genes and for proteins encoded thereby, and thus enables the practitioner to assess, predict, and affect the physiological state of various human and murine tissues.

SUMMARY OF THE INVENTION

The present invention is based, at least in part, on the discovery of a variety of human and murine cDNA molecules which encode proteins which are herein designated TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. These seven proteins, fragments thereof, derivatives thereof, and variants thereof are collectively referred to herein as the polypeptides of the invention or the proteins of the invention. Nucleic acid molecules encoding polypeptides of the invention are collectively referred to as nucleic acids of the invention.

The nucleic acids and polypeptides of the present invention are useful as modulating agents in regulating a variety of cellular processes. Accordingly, in one aspect, the present invention provides isolated nucleic acid molecules encoding a polypeptide of the invention or a biologically active portion thereof. The present invention also provides nucleic acid molecules which are suitable as primers or hybridization probes for the detection of nucleic acids encoding a polypeptide of the invention.

The invention also features nucleic acid molecules which are at least 40% (or 50%, 60%, 70%, 80%, 90%, 95%, or 98%) identical to the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of a cDNA clone deposited with ATCC® as one of Accession numbers 207219, 207184, 207228, 207185, 207220, and 207221 (“a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221”), or a complement thereof.

The invention features nucleic acid molecules which include a fragment of at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300, 350, 400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3500, 4000, 4500, or 4928) consecutive nucleotide residues of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, or a complement thereof.

The invention also features nucleic acid molecules which include a nucleotide sequence encoding a protein having an amino acid sequence that is at least 50% (or 60%, 70%, 80%, 90%, 95%, or 98%) identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, or a complement thereof.

In preferred embodiments, the nucleic acid molecules have the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or the nucleotide sequence of a cDNA of a clone deposited as one of ATCC4 207219, 207184, 207228, 207185, 207220, and 207221.

Also within the invention are nucleic acid molecules which encode a fragment of a polypeptide having the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, the fragment including at least 8 (10, 15, 20, 25, 30, 40, 50, 75, 100, 125, 150, or 200) consecutive amino acids of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221.

The invention includes nucleic acid molecules which encode a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, wherein the nucleic acid molecule hybridizes under stringent conditions to a nucleic acid molecule having a nucleic acid sequence encoding any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, or a complement thereof.

Also within the invention are isolated polypeptides or proteins having an amino acid sequence that is at least about 50%, preferably 60%, 75%, 90%, 95%, or 98% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74.

Also within the invention are isolated polypeptides or proteins which are encoded by a nucleic acid molecule having a nucleotide sequence that is at least about 40%, preferably 50%, 75%, 85%, or 95% identical the nucleic acid sequence encoding any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, and isolated polypeptides or proteins which are encoded by a nucleic acid molecule consisting of the nucleotide sequence which hybridizes under stringent hybridization conditions to a nucleic acid molecule having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73.

Also within the invention are polypeptides which are naturally occurring allelic variants of a polypeptide that includes the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes under stringent conditions to a nucleic acid molecule having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a complement thereof.

The invention also features nucleic acid molecules that hybridize under stringent conditions to a nucleic acid molecule having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of a cDNA of a clone deposited as one of ATTC® 207219, 207184, 207228, 207185, 207220, and 207221, or a complement thereof. In other embodiments, the nucleic acid molecules are at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300, 350, 400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3500, 4000, 4500, or 4928) nucleotides in length and hybridize under stringent conditions to a nucleic acid molecule having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221, or a complement thereof. In some embodiments, the isolated nucleic acid molecules encode a cytoplasmic, transmembrane, extracellular, or other domain of a polypeptide of the invention. In other embodiments, the invention provides an isolated nucleic acid molecule which is antisense to the coding strand of a nucleic acid of the invention.

Another aspect of the invention provides vectors, e.g., recombinant expression vectors, comprising a nucleic acid molecule of the invention. In another embodiment, the invention provides isolated host cells, e.g., mammalian and non-mammalian cells, containing such a vector or a nucleic acid of the invention. The invention also provides methods for producing a polypeptide of the invention by culturing, in a suitable medium, a host cell of the invention containing a recombinant expression vector encoding a polypeptide of the invention such that the polypeptide of the invention is produced.

Another aspect of this invention features isolated or recombinant proteins and polypeptides of the invention. Preferred proteins and polypeptides possess at least one biological activity possessed by the corresponding naturally-occurring human polypeptide. An activity, a biological activity, and a functional activity of a polypeptide of the invention refers to an activity exerted by a protein or polypeptide of the invention on a responsive cell as determined in vivo, or in vitro, according to statndard techniques.

Such activities can be a direct activity, such as an association with or an association with or an enzymatic activity on a second protein, or an indirect activity, such as a cellular process (e.g., signaling activity) mediated by interaction of the protein with a second protein. Such activities include, by way of example, formation of protein-protein interactions with proteins of one or more signaling pathways (e.g., with a protein with which the naturally-occuring polypeptide interacts); binding with a ligand of the naturally-occuring protein; and binding with an intracellular target of the naturally-occuring protein. Other activities include modulation of one or more of cellular proliferation, of cellular differentiation, of chemotaxis, of cellular migration, and of cell death (e.g., apoptosis).

By way of example, TANGO 202 exhibits the ability to affect growth, proliferation, survival, differentiation, and activity of human hematopoietic cells (e.g., bone marrow stromal cells) and fetal cells. TANGO 202 modulates cellular binding to one or more mediators, modulates proteolytic activity in vivo, modulates developmental processes, and modulates cell growth, proliferation, survival, differentiation, and activity. Thus, TANGO 202 can be used to prevent, diagnose, or treat disorders relating to aberrant cellular protease activity, inappropriate interaction (or non-interaction) of cells with mediators, inappropriate development, and blood and hematopoietic cell-related disorders. Exemplary disorders for which TANGO 202 is useful include immune disorders, infectious diseases, auto-immune disorders, vascular and cardiovascular disorders, disorders related to mal-expression of growth factors, cancers, hematological disorders, various cancers, birth defects, developmental defects, and the like.

Further by way of example, TANGO 234 exhibits the ability to affect growth, proliferation, survival, differentiation, and activity of human lung, hematopoietic, and fetal cells and of (e.g., bacterial or fungal) cells and viruses which infect humans. TANGO 234 modulates growth, proliferation, survival, differentiation, and activity of gamma delta T cells, for example. Furthermore, TANGO 234 modulates cholesterol deposition on human arterial walls, and is involved in uptake and metabolism of low density lipoprotein and regulation of serum cholesterol levels. Thus, TANGO 234 can be used to affect development and persistence of atherogenesis and arteriosclerosis, as well as other vascular and cardiovascular disorders. Other exemplary disorders for which TANGO 234 is useful include immune development disorders and disorders involving generation and persistence of an immune response to bacterial, fungal, and viral infections.

Still further by way of example, TANGO 265 modulates growth and regeneration of neuronal and epithelial tissues, and guides neuronal axon development. TANGO 265 is a transmembrane protein which mediates cellular interaction with cells, molecules and structures (e.g., extracellular matrix) in the extracellular environment. TANGO 265 is therefore involved in growth, organization, and adhesion of tissues and the cells which constitute those tissues. Furthermore, TANGO 265 modulates growth, proliferation, survival, differentiation, and activity of neuronal cells and immune system cells. Thus, TANGO 265 can be used, for example, to prevent, diagnose, or treat disorders characterized by aberrant organization or development of a tissue or organ, for guiding neural axon development, for modulating differentiation of cells of the immune system, for modulating cytokine production by cells of the immune system, for modulating reactivity of cells of the immune system toward cytokines, for modulating initiation and persistence of an inflammatory response, and for modulating proliferation of epithelial cells.

Yet further by way of example, TANGO 273 protein mediates one or more physiological responses of cells to bacterial infection, e.g., by mediating one or more of detection of bacteria in a tissue in which it is expressed, movement of cells with relation to sites of bacterial infection, production of biological molecules which inhibit bacterial infection, and production of biological molecules which alleviate cellular or other physiological damage wrought by bacterial infection. TANGO 273, a transmembrane protein, is also involved in transmembrane signal transduction, and therefore mediates transmission of signals between the extracellular and intracellular environments of cells. TANGO 273 mediates regulation of cell growth and proliferation, endocytosis, activation of respiratory burst, and other physiological processes triggered by transmission of a signal via a protein with which TANGO 273 interacts. The compositions and methods of the invention can therefore be used to prevent, diagnose, and treat disorders involving one or more physiological activities mediated by TANGO 273 protein. Such disorders include, for example, various bone-related disorders such as metabolic, homeostatic, and developmental bone disorders (e.g., osteoporosis, various cancers, skeletal development disorders, bone fragility and the like), disorders caused by or related to bacterial infection, and disorders characterized by aberrant transmembrane signal transduction by TANGO 273.

As an additional example, TANGO 286 protein is involved in lipid-binding physiological processes such as lipid transport, metabolism, serum lipid particle regulation, host anti-microbial defensive mechanisms, and the like. Thus, the compositions and methods of the invention can therefore be used to prevent, diagnose, and treat disorders involving one or more physiological activities mediated by TANGO 286 protein. Such disorders include, for example, lipid transport disorders, lipid metabolism disorders, obesity, disorders of serum lipid particle regulation, disorders involving insufficient or inappropriate host anti-microbial defensive mechanisms, vasculitis, bronchiectasis, LPS-related disorders such as shock, disseminated intravascular coagulation, anemia, thrombocytopenia, adult respiratory distress syndrome, renal failure, liver disease, and disorders associated with Gran negative bacterial infections, such as bacteremia, endotoxemia, sepsis, and the like.

Further by way of example, TANGO 294 protein is involved in facilitating absorption and metabolism of fat. Thus, the compositions and methods of the invention can therefore be used to prevent, diagnose, and treat disorders involving one or more physiological activities mediated by TANGO 294 protein. Such disorders include, for example, inadequate expression of gastric/pancreatic lipase, cystic fibrosis, exocrine pancreatic insufficiency, medical treatments which alter fat absorption, obesity, and the like.

As another example, INTERCEPT 296 protein is involved in physiological processes related to disorders of the human lung and esophagus. Thus, the compositions and methods of the invention can be used to prevent, diagnose, and treat these disorders. Such disorders include, for example, various cancers, bronchitis, cystic fibrosis, respiratory infections (e.g., influenza, bronchiolitis, pneumonia, and tuberculosis), asthma, emphysema, chronic bronchitis, bronchiectasis, pulmonary edema, pleural effusion, pulmonary embolus, adult and infant respiratory distress syndromes, heartburn, and gastric reflux esophageal disease.

In one embodiment, a polypeptide of the invention has an amino acid sequence sufficiently identical to an identified domain of a polypeptide of the invention. As used herein, the term “sufficiently identical” refers to a first amino acid or nucleotide sequence which contains a sufficient or minimum number of identical or equivalent (e.g., with a similar side chain) amino acid residues or nucleotides to a second amino acid or nucleotide sequence such that the first and second amino acid or nucleotide sequences have a common structural domain and/or common functional activity. For example, amino acid or nucleotide sequences which contain a common structural domain having about 65% identity, preferably 75% identity, more preferably 85%, 95%, or 98% identity are defined herein as sufficiently identical.

In one embodiment, the isolated polypeptide of the invention lacks both a transmembrane and a cytoplasmic domain. In another embodiment, the polypeptide lacks both a transmembrane domain and a cytoplasmic domain and is soluble under physiological conditions.

The polypeptides of the present invention, or biologically active portions thereof, can be operably linked to a heterologous amino acid sequence to form fusion proteins. The invention further features antibody substances that specifically bind a polypeptide of the invention such as monoclonal or polyclonal antibodies, antibody fragments, single-chain antibodies, and the like. In addition, the polypeptides of the invention or biologically active portions thereof can be incorporated into pharmaceutical compositions, which optionally include pharmaceutically acceptable carriers. These antibody substances can be made, for example, by providing the polypeptide of the invention to an immunocompetent vertebrate and thereafter harvesting blood or serum from the vertebrate.

In another aspect, the present invention provides methods for detecting the presence of the activity or expression of a polypeptide of the invention in a biological sample by contacting the biological sample with an agent capable of detecting an indicator of activity such that the presence of activity is detected in the biological sample.

In another aspect, the invention provides methods for modulating activity of a polypeptide of the invention comprising contacting a cell with an agent that modulates (inhibits or enhances) the activity or expression of a polypeptide of the invention such that activity or expression in the cell is modulated. In one embodiment, the agent is an antibody that specifically binds to a polypeptide of the invention.

In another embodiment, the agent modulates expression of a polypeptide of the invention by modulating transcription, splicing, or translation of an mRNA encoding a polypeptide of the invention. In yet another embodiment, the agent is a nucleic acid molecule having a nucleotide sequence that is antisense with respect to the coding strand of an mRNA encoding a polypeptide of the invention.

The present invention also provides methods to treat a subject having a disorder characterized by aberrant activity of a polypeptide of the invention or aberrant expression of a nucleic acid of the invention by administering an agent which is a modulator of the activity of a polypeptide of the invention or a modulator of the expression of a nucleic acid of the invention to the subject. In one embodiment, the modulator is a protein of the invention. In another embodiment, the modulator is a nucleic acid of the invention. In other embodiments, the modulator is a peptide, peptidomimetic, or other small molecule (e.g., a small organic molecule).

The present invention also provides diagnostic assays for identifying the presence or absence of a genetic lesion or mutation characterized by at least one of: (i) aberrant modification or mutation of a gene encoding a polypeptide of the invention, (ii) mis-regulation of a gene encoding a polypeptide of the invention, and (iii) aberrant post-translational modification of a polypeptide of the invention wherein a wild-type form of the gene encodes a polypeptide having the activity of the polypeptide of the invention.

In another aspect, the invention provides a method for identifying a compound that binds to or modulates the activity of a polypeptide of the invention. In general, such methods entail measuring a biological activity of the polypeptide in the presence and absence of a test compound and identifying those compounds which alter the activity of the polypeptide.

The invention also features methods for identifying a compound which modulates the expression of a polypeptide or nucleic acid of the invention by measuring the expression of the polypeptide or nucleic acid in the presence and absence of the compound.

In yet a further aspect, the invention provides substantially purified antibodies or fragments thereof (i.e., antibody substances), including non-human antibodies or fragments thereof, which specifically bind with a polypeptide of the invention or with a portion thereof. In various embodiments, these substantially purified antibodies/fragments can be human, non-human, chimeric, and/or humanized antibodies. Non-human antibodies included in the invention include, by way of example, goat, mouse, sheep, horse, chicken, rabbit, and rat antibodies. In addition, the antibodies of the invention can be polyclonal antibodies or monoclonal antibodies.

In a particularly preferred embodiment, the antibody substance of the invention specifically binds with an extracellular domain of one of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. Preferably, the extracellular domain with which the antibody substance binds has an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 14, 22, 30, 37, 49, 50, and 56-58.

Any of the antibody substances of the invention can be conjugated with a therapeutic moiety or with a detectable substance. Non-limiting examples of detectable substances that can be conjugated with the antibody substances of the invention include an enzyme, a prosthetic group, a fluorescent material (i.e., a fluorophore), a luminescent material, a bioluminescent material, and a radioactive material (e.g., a radionuclide or a substituent comprising a radionuclide).

The invention also provides a kit containing an antibody substance of the invention conjugated with a detectable substance, and instructions for use. Still another aspect of the invention is a pharmaceutical composition comprising an antibody substance of the invention and a pharmaceutically acceptable carrier. In preferred embodiments, the pharmaceutical composition contains an antibody substance of the invention, a therapeutic moiety (preferably conjugated with the antibody substance), and a pharmaceutically acceptable carrier.

Other features and advantages of the invention will be apparent from the following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 comprises FIGS. 1A-1M. The nucleotide sequence (SEQ ID NO: 1) of a cDNA encoding the human TANGO 202 protein described herein is listed in FIGS. 1A-1D. The open reading frame (ORF; residues 34 to 1458; SEQ ID NO: 2) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 3) of human TANGO 202 is listed. The nucleotide sequence (SEQ ID NO: 67) of a cDNA encoding the murine TANGO 202 protein described herein is listed in FIGS. 1E-1I. The ORF (residues 81 to 1490; SEQ ID NO: 68) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 69) of murine TANGO 202 is listed. An alignment of the amino acid sequences of human (“Hum.”; SEQ ID NO: 3) and murine (“Mur.”; SEQ ID NO: 69) TANGO 202 protein is shown in FIGS. 1J and 1K, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. FIG. 1L is a hydrophilicity plot of human TANGO 202 protein, in which the locations of cysteine residues (“Cys”) and potential N-glycosylation sites (“Ngly”) are indicated by vertical bars and the predicted extracellular (“out”), intracellular (“ins”), or transmembrane (“TM”) locations of the protein backbone is indicated by a horizontal bar. FIG. 1M is a hydrophilicity plot of murine TANGO 202 protein.

FIG. 2 comprises FIGS. 2A-2Qxvii. The nucleotide sequence (SEQ ID NO: 9) of a cDNA encoding the human TANGO 234 protein described herein is listed in FIGS. 2A-2I. The ORF (residues 28 to 4386; SEQ ID NO: 10) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 11) of human TANGO 234 is listed. FIG. 2J is a hydrophilicity plot of human TANGO 234 protein. An alignment of the amino acid sequences of human TANGO 234 (“Hum”; SEQ ID NO: 11) and bovine WC1 (“WC1”; SEQ ID NO: 78) proteins is shown in FIGS. 2K-2P, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. An alignment of the nucleotide sequences of an ORF encoding human TANGO 234 (“Hum”; SEQ ID NO: 10) and an ORF encoding bovine WC1 (“WC1”; SEQ ID NO: 79) proteins is shown in FIGS. 2Qi-2Qxvii, wherein identical nucleotide residues are indicated by “:”.

FIG. 3 comprises FIGS. 3A-3U. The nucleotide sequence (SEQ ID NO: 17) of a cDNA encoding the human TANGO 265 protein described herein is listed in FIGS. 3A-3E. The ORF (residues 32 to 2314; SEQ ID NO: 18) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 19) of human TANGO 265 is listed. An alignment of the amino acid sequences of human TANGO 265 protein (“Hum.”; SEQ ID NO: 19) and murine semaphorin B protein (“Mur.”; SEQ ID NO: 70; GenBank Accession No. X85991) is shown in FIGS. 3F-3H, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. In FIGS. 3I-3T, an alignment of the nucleotide sequences of the cDNA encoding human TANGO 265 protein (“Hum.”; SEQ ID NO: 17) and the nucleotide sequences of the cDNA encoding murine semaphorin B protein (“Mur.”; SEQ ID NO: 71; GenBank Accession No. X85991) is shown. FIG. 3U is a hydrophilicity plot of TANGO 265 protein.

FIG. 4 comprises FIGS. 4A-4J. The nucleotide sequence (SEQ ID NO: 25) of a cDNA encoding the human TANGO 273 protein described herein is listed in FIGS. 4A-4C. The ORF (residues 135 to 650; SEQ ID NO: 26) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 27) of human TANGO 273 is listed. The nucleotide sequence (SEQ ID NO: 72) of a cDNA encoding the murine TANGO 273 protein described herein is listed in FIGS. 4D-4G. The ORF (residues 137 to 652; SEQ ID NO: 73) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 74) of murine TANGO 273 is listed. An alignment of the amino acid sequences of human (“Hum.”; SEQ ID NO: 27) and murine (“Mur.”; SEQ ID NO: 74) TANGO 273 protein is shown in FIG. 4H, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. FIG. 4I is a hydrophilicity plot of human TANGO 273 protein, and FIG. 4J is a hydrophilicity plot of murine TANGO 273 protein.

FIG. 5 comprises FIGS. 5A-5I. The nucleotide sequence (SEQ ID NO: 33) of a cDNA encoding the human TANGO 286 protein described herein is listed in FIGS. 5A-5D. The ORF (residues 133 to 1497; SEQ ID NO: 34) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 35) of human TANGO 286 is listed. FIG. 5E is a hydrophilicity plot of TANGO 286 protein. An alignment of the amino acid sequences of human TANGO 286 (“286”; SEQ ID NO: 35) and BPI protein (“BPI”; SEQ ID NO: 38) protein is shown in FIGS. 5F and 5G, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. An alignment of the amino acid sequences of human TANGO 286 (“286”; SEQ ID NO: 35) and RENP protein (“RENP”; SEQ ID NO: 39) is shown in FIGS. 5H and 5I, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”.

FIG. 6 comprises FIGS. 6A-6H. The nucleotide sequence (SEQ ID NO: 45) of a cDNA encoding the human TANGO 294 protein described herein is listed in FIGS. 6A-6C. The ORF (residues 126 to 1394; SEQ ID NO: 46) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 47) of human TANGO 294 is listed. An alignment of the amino acid sequences of human TANGO 294 protein (“294”; SEQ ID NO: 47) and a known human lipase protein (“HLP”; SEQ ID NO: 75; GenBank Accession No. NP_(—)004181) is shown in FIGS. 6D and 6E, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”. FIG. 6F is a hydrophilicity plot of TANGO 294 protein. An alignment of the amino acid sequences of human TANGO 294 protein (“294”; SEQ ID NO: 47) and a known human lysosomal acid lipase protein (“LAL”; SEQ ID NO: 41) is shown in FIGS. 6G and 6H, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”.

FIG. 7 comprises FIGS. 7A-7F. The nucleotide sequence (SEQ ID NO: 53) of a cDNA encoding the human INTERCEPT 296 protein described herein is At listed in FIGS. 7A-7C. The ORF (residues 70 to 1098; SEQ ID NO: 54) of the cDNA is indicated by nucleotide triplets, above which the amino acid sequence (SEQ ID NO: 55) of human INTERCEPT 296 protein is listed. FIG. 7D is a hydrophilicity plot of INTERCEPT 296 protein. An alignment of the amino acid sequences of human INTERCEPT 296 protein (“296”; SEQ ID NO: 55) and C. elegans C06E1.3 related protein (“CRP”; SEQ ID NO: 40) is shown in FIGS. 7E and 7F, wherein identical amino acid residues are indicated by “:” and similar amino acid residues are indicated by “.”.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, at least in part, on the discovery of a variety of human and murine cDNA molecules which encode proteins which are herein designated TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. These proteins exhibit a variety of physiological activities, and are included in a single application for the sake of convenience. It is understood that the allowability or non-allowability of claims directed to one of these proteins has no bearing on the allowability of claims directed to the others. The characteristics of each of these proteins and the cDNAs encoding them are now described separately.

TANGO 202

A cDNA clone (designated jthke096b05) encoding at least a portion of human TANGO 202 protein was isolated from a human fetal skin cDNA library. The corresponding murine cDNA was isolated as a clone (designated jtmMa044f07) from a bone marrow stromal cell cDNA library. The human TANGO 202 protein is predicted by structural analysis to be a type I membrane protein, although it can exist in a secreted form as well. The murine TANGO 202 protein is predicted by structural analysis to be a secreted protein.

The full length of the cDNA encoding human TANGO 202 protein (FIG. 1; SEQ ID NO: 1) is 1656 nucleotide residues. The open reading frame (ORF) of this cDNA, nucleotide residues 34 to 1458 of SEQ ID NO: 1 (i.e., SEQ ID NO: 2), encodes a 475-amino acid transmembrane protein (FIG. 1; SEQ ID NO: 3).

The invention thus includes purified human TANGO 202 protein, both in the form of the immature 475 amino acid residue protein (SEQ ID NO: 3) and in the form of the mature 456 amino acid residue protein (SEQ ID NO: 5). The invention also includes purified murine TANGO 202 protein, both in the form of the immature 470 amino acid residue protein (SEQ ID NO: 67) and in the form of the mature 451 amino acid residue protein (SEQ ID NO: 43). Mature human or murine TANGO 202 proteins can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or they can be synthesized by generating immature TANGO 202 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature human and murine TANGO 202 proteins, the invention includes fragments, derivatives, and variants of these TANGO 202 proteins, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 1 or some portion thereof or SEQ ID NO: 67 or some portion thereof, such as the portion which encodes mature human or murine TANGO 202 protein, immature human or murine TANGO 202 protein, or a domain of human or murine TANGO 202 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 202 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features. As used herein, the term “family” is intended to mean two or more proteins or nucleic acid molecules having a common or similar domain structure and having sufficient amino acid or nucleotide sequence identity as defined herein. Family members can be from either the same or different species (e.g., human and mouse, as described herein). For example, a family can comprise two or more proteins of human origin, or can comprise one or more proteins of human origin and one or more of non-human origin.

A common domain present in TANGO 202 proteins is a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound and secreted proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 202 protein contains a signal sequence corresponding to amino acid residues 1 to 19 of SEQ ID NO: 3 (SEQ ID NO: 4) or to amino acid residues 1 to 19 of SEQ ID NO: 69 (SEQ ID NO: 42). The signal sequence is cleaved during processing of the mature protein.

TANGO 202 proteins can also include an extracellular domain. As used herein, an “extracellular domain” refers to a portion of a protein which is localized to the non-cytoplasmic side of a lipid bilayer of a cell when a nucleic acid encoding the protein is expressed in the cell. The human TANGO 202 protein extracellular domain is located from about amino acid residue 20 to about amino acid residue 392 of SEQ ID NO: 3 in the non-secreted form, and from about amino acid residue 20 to amino acid residue 475 of SEQ ID NO: 3 (i.e., the entire mature human protein). The murine TANGO 202 protein extracellular domain is located from about amino acid residue 20 to amino acid residue 470 of SEQ ID NO: 69 (i.e., the entire mature murine protein).

TANGO 202 proteins of the invention can also include a transmembrane domain. As used herein, a “transmembrane domain” refers to an amino acid sequence having at least about 20 to 25 amino acid residues in length and which contains at least about 65-70% hydrophobic amino acid residues such as alanine, leucine, phenylalanine, protein, tyrosine, tryptophan, or valine. In a preferred embodiment, a transmembrane domain contains at least about 15 to 30 amino acid residues, preferably about 20-25 amino acid residues, and has at least about 60-80%, more preferably 65-75%, and more preferably at least about 70% hydrophobic residues. Thus, in one embodiment, a TANGO 202 protein of the invention contains a transmembrane domain corresponding to about amino acid residues 393 to 415 of SEQ ID NO: 3 (SEQ ID NO: 7).

In addition, TANGO 202 proteins of the invention can include a cytoplasmic domain, particularly including a carboxyl-terminal cytoplasmic domain. As used herein, a “cytoplasmic domain” refers to a portion of a protein which is localized to the cytoplasmic side of a lipid bilayer of a cell when a nucleic acid encoding the protein is expressed in the cell. The cytoplasmic domain is located from about amino acid residue 416 to amino acid residue 475 of SEQ ID NO: 3 (SEQ ID NO: 8) in the non-secreted form of human TANGO 202 protein.

TANGO 202 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Tables I (for human TANGO 202) and II (for murine TANGO 202), as predicted by computerized sequence analysis of TANGO 202 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 202 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}).

TABLE I Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 3 Sequence N-glycosylation site 47 to 50 NWTA 61 to 64 NETF 219 to 222 NYSA 295 to 298 NVSL 335 to 338 NQTV 347 to 350 NLSV Protein kinase C 70 to 72 TLK phosphorylation site 137 to 139 TSK 141 to 143 SNK 155 to 157 SQR 238 to 240 TGR 245 to 247 TIR 277 to 279 THR 307 to 309 SDR 355 to 357 SSK 387 to 389 SHR 418 to 420 TFK 421 to 423 SHR Casein kinase II phosphorylation 337 to 340 TVAE site 438 to 441 TSGE 464 to 467 SQQD N-myristoylation site 53 to 58 GGKPCL 120 to 125 GNLGCY 136 to 141 GTSKTS 162 to 167 GMESGY 214 to 219 GACGGN Kringle domain signature 85 to 90 YCRNPD Kringle Domain  34 to 116 See FIG. 1 CUB domain 216 to 320 See FIG. 1

TABLE II Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 69 Sequence N-glycosylation site 59 to 62 NETF 217 to 220 NYSA 255 to 258 NFTL 293 to 296 NVSL 333 to 336 NQTL 345 to 348 NLSV cAMP- or cGMP-dependent 455 to 458 RRSS protein kinase phosphorylation site Protein kinase C phosphorylation 68 to 70 TLK site 135 to 137 TSK 139 to 141 SNK 153 to 155 SQR 236 to 238 TGR 243 to 245 TIR 275 to 277 THR 283 to 285 SGR 305 to 307 SDR 353 to 355 SSK 408 to 410 SQR 453 to 455 SLR 457 to 459 SSR Casein kinase II 28 to 31 SGPE phosphorylation site 257 to 260 TLFD 321 to 324 TKEE 335 to 338 TLAE 384 to 387 TATE N-myristoylation site 51 TO 56 GGKPCL 118 TO 123 GNLGCY 134 TO 139 GTSKTS 160 TO 165 GMESGY 212 TO 217 GACGGN 391 TO 396 GLCTAW 429 TO 434 GTVVSL Kringle domain signature 83 to 88 YCRNPD Kringle Domain  32 to 114 See FIG. 1 CUB domain 214 to 318 See FIG. 1

As used herein, the term “post-translational modification site” refers to a protein domain that includes about 3 to 10 amino acid residues, more preferably about 3 to 6 amino acid residues wherein the domain has an amino acid sequence which comprises a consensus sequence which is recognized and modified by a protein-modifying enzyme. Exemplary protein-modifying enzymes include amino acid glycosylases, cAMP- and cGMP-dependent protein kinases, protein kinase C, casein kinase II, myristoylases, and prenyl transferases. In various embodiments, the protein of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational modification sites described herein in Tables I and II.

Exemplary additional domains present in human and murine TANGO 202 protein include Kringle domains and CUB domains. In one embodiment, the protein of the invention has at least one domain that is at least 55%, preferably at least about 65%, more preferably at least about 75%, yet more preferably at least about 85%, and most preferably at least about 95% identical to one of the domains described herein in Tables I and II. Preferably, the protein of the invention has at least one Kringle domain and one CUB domain.

A Kringle domain has a characteristic profile that has been described in the art (Castellino and Beals (1987) J. Mol. Evol. 26:358-369; Patthy (1985) Cell 41:657-663; Ikeo et al. (1991) FEBS Lett. 287:146-148). Many, but not all, Kringle domains comprise a conserved hexapeptide signature sequence, namely

(F or Y)-C-R-N-P-(D or N or R).

The cysteine residue is involved in a disulfide bond.

Kringle domains are triple-looped, disulfide cross-linked domains found in a varying number of copies in, for example, some serine proteases and plasma proteins. Kringle domains have a role in binding mediators (e.g., membranes, other proteins, or phospholipids) and in regulation of proteolytic activity. Kringle domains have been identified in the following proteins, for example: apolipoprotein A, blood coagulation factor XII (Hageman factor), hepatocyte growth factor (HGF), HGF-like protein (Friezner Degen et al., (1991) Biochemistry 30:9781-9791), HGF activator (Miyazawa et al., (1993) J. Biol. Chem. 268:10024-10028), plasminogen, thrombin, tissue plasminogen activator, urokinase-type plasminogen activator, and four influenza neuraminidases. The presence of a Kringle domain in each of human and murine TANGO 202 protein indicates that TANGO 202 is involved in one or more physiological processes in which these other Kringle domain-containing proteins are involved, has biological activity in common with one or more of these other Kringle domain-containing proteins, or both.

CUB domains are extracellular domains of about 110 amino acid residues which occur in functionally diverse, mostly developmentally regulated proteins (Bork and Beckmann (1993) J. Mol. Biol. 231:539-545; Bork (1991) FEBS Lett. 282:9-12). Many CUB domains contain four conserved cysteine residues, although some, like that of TANGO 202, contain only two of the conserved cysteine residues. The structure of the CUB domain has been predicted to assume a beta-barrel configuration, similar to that of immunoglobulins. Other proteins which have been found to comprise one or more CUB domains include, for example, mammalian complement sub-components Cls and Clr, hamster serine protease Casp, mammalian complement activating component of Ra-reactive factor, vertebrate enteropeptidase, vertebrate bone morphogenic protein 1, sea urchin blastula proteins BP10 and SpAN, Caenorhabditis elegans hypothetical proteins F42A10.8 and R151.5, neuropilin (A5 antigen), sea urchin fibropellins I and III, mammalian hyaluronate-binding protein TSG6 (PS4), mammalian spermadhesins, and Xenopus embryonic protein UVS.2. The presence of a CUB domain in each of human and murine TANGO 202 protein indicates that TANGO 202 is involved in one or more physiological processes in which these other CUB domain-containing proteins are involved, has biological activity in common with one or more of these other CUB domain-containing proteins, or both.

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that human TANGO 202 protein includes a 19 amino acid signal peptide (amino acid residues 1 to 19 of SEQ ID NO: 3; SEQ ID NO: 4) preceding the mature TANGO 202 protein (amino acid residues 20 to 475 of SEQ ID NO: 3; SEQ ID NO: 5). Human TANGO 202 protein includes an extracellular domain (amino acid residues 20 to 392 of SEQ ID NO: 3; SEQ ID NO: 6); a transmembrane domain (amino acid residues 393 to 415 of SEQ ID NO: 3; SEQ ID NO: 7); and a cytoplasmic domain (amino acid residues 416 to 475 of SEQ ID NO: 3; SEQ ID NO: 8). The murine homolog of TANGO 202 protein is predicted to be a secreted protein. Thus, it is recognized that human TANGO 202 can also exist in the to form of a secreted protein, likely being translated from an alternatively spliced TANGO 202 mRNA. In a variant form of the protein, an extracellular portion of TANGO 202 protein (e.g., amino acid residues 20 to 392 of SEQ ID NO: 3) can be cleaved from the mature protein to generate a soluble fragment of TANGO 202.

FIG. 1L depicts a hydrophilicity plot of human TANGO 202 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 19 of SEQ ID NO: 3 is the signal sequence of human TANGO 202 (SEQ ID NO: 4). The hydrophobic region which corresponds to amino acid residues 393 to 415 of SEQ ID NO: 3 is the transmembrane domain of human TANGO 202 (SEQ ID NO: 7). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 202 protein from about amino acid residue 61 to about amino acid residue 95 appears to be located at or near the surface of the protein, while the region from about amino acid residue 395 to about amino acid residue 420 appears not to be located at or near the surface.

The predicted molecular weight of human TANGO 202 protein without modification and prior to cleavage of the signal sequence is about 51.9 kilodaltons. The predicted molecular weight of the mature human TANGO 202 protein without modification and after cleavage of the signal sequence is about 50.1 kilodaltons.

The full length of the cDNA encoding murine TANGO 202 protein (FIG. 1; SEQ ID NO: 67) is 4928 nucleotide residues. The ORF of this cDNA, nucleotide residues 81 to 1490 of SEQ ID NO: 67 (i.e., SEQ ID NO: 68), encodes a 470-amino acid secreted protein (FIG. 1; SEQ ID NO: 69).

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that murine TANGO 202 protein includes a 19 amino acid signal peptide (amino acid residues 1 to 19 of SEQ ID NO: 69; SEQ ID NO: 42) preceding the mature TANGO 202 protein (amino acid residues 20 to 470 of SEQ ID NO: 69; SEQ ID NO: 43). Murine TANGO 202 protein is a secreted protein.

FIG. 1M depicts a hydrophilicity plot of murine TANGO 202 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 19 of SEQ ID NO: 69 is the signal sequence of murine TANGO 202 (SEQ ID NO: 42). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of murine TANGO 202 protein from about amino acid residue 61 to about amino acid residue 95 appears to be located at or near the surface of the protein, while the region from about amino acid residue 295 to about amino acid residue 305 appears not to be located at or near the surface

The predicted molecular weight of murine TANGO 202 protein without modification and prior to cleavage of the signal sequence is about 51.5 kilodaltons. The predicted molecular weight of the mature murine TANGO 202 protein without modification and after cleavage of the signal sequence is about 49.7 kilodaltons.

Human and murine TANGO 202 proteins exhibit considerable sequence similarity, as indicated herein in FIGS. 1J and 1K. FIGS. 1J and 1K depict an alignment of human and murine TANGO 202 amino acid sequences (SEQ ID NOs: 3 and 69, respectively). In this alignment (made using the ALIGN software {Myers and Miller (1989) CABIOS, ver. 2.0}; pam120.mat scoring matrix; gap penalties −12/−4), the proteins are 76.5% identical. The human and murine ORFs encoding TANGO 202 are 87.4% identical, as assessed using the same software and parameters.

In situ hybridization experiments in mouse tissues indicated that mRNA corresponding to the cDNA encoding TANGO 202 is expressed in the tissues listed in Table III, wherein “+” indicates detectable expression and “++” indicates a greater level of expression than “+”.

TABLE III Relative Level of Animal Tissue Expression Mouse bladder, especially in ++ (Adult) transitional epithelium renal glomeruli + brain + heart + liver + spleen + placenta + Mouse ubiquitous + (Embryo)

Biological Function of TANGO 202 Proteins, Nucleic Acids, and Modulators Thereof

TANGO 202 proteins are involved in disorders which affect both tissues in which they are normally expressed and tissues in which they are normally not expressed. Based on the observation that TANGO 202 is expressed in human fetal skin, ubiquitously in fetal mouse tissues, in adult murine bone marrow stromal cells, and in cells of adult murine bladder, renal glomeruli, brain, heart, liver, spleen and placenta, TANGO 202 protein is involved in one or more biological processes which occur in these tissues. In particular, TANGO 202 is involved in modulating growth, proliferation, survival, differentiation, and activity of cells of these tissues including, but not limited to, hematopoietic and fetal cells. Thus, TANGO 202 has a role in disorders which affect these cells and their growth, proliferation, survival, differentiation, and activity. Ubiquitous expression of TANGO 202 in fetal murine tissues, contrasted with limited expression in adult murine tissues further indicates that TANGO 202 is involved in disorders in which it is inappropriately expressed (e.g., disorders in which TANGO 202 is expressed in adult murine tissues other than bone marrow stromal cells and disorders in which TANGO 202 is not expressed in one or more developing fetal tissues).

The presence of a Kringle domain in both the murine and human TANGO 202 proteins indicates that this protein is involved in modulating cellular binding to one or more mediators (e.g., proteins, phospholipids, intracellular organelles, or other cells), in modulating proteolytic activity, or both. The presence of a Kringle domain in other proteins (e.g., growth factors) indicates activities that these proteins share with TANGO 202 protein (e.g., modulating cell dissociation and migration into and through extracellular matrices). The presence of Kringle domains in numerous plasma proteins, particularly coupled with the observation that TANGO 202 is expressed in adult murine bone marrow stromal cells, indicates a role for TANGO 202 protein in modulating binding of blood or hematopoietic cells (or both) to one or more mediators. Thus, TANGO 202 is involved in disorders relating to aberrant cellular protease activity, inappropriate interaction or non-interaction of cells with mediators, and in blood and hematopoietic cell-related disorders. Such disorders include, by way of example and not limitation, immune disorders, infectious diseases, auto-immune disorders, vascular and cardiovascular disorders, disorders related to mal-expression of growth factors, cancers, hematological disorders, and the like.

The cDNA encoding TANGO 202 exhibits significant nucleotide sequence similarity with a polynucleotide encoding a kringle-domain-containing protein (designated HTHBZ47) described in the European Patent Application No. EP 0 911 399 A2 (published Apr. 28, 1999). Thus, the TANGO 202 protein can exhibit one or more of the activities exhibited by HTHBZ47, and can be used to prevent, inhibit, diagnose, and treat one or more disorders for which HTHBZ47 is useful. These disorders include cancer, inflammation, autoimmune disorders, allergic disorders, asthma, rheumatoid arthritis, inflammation of central nervous system tissues, cerebellar degeneration, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amylotrophic lateral sclerosis, head injury damage and other neurological abnormalities, septic shock, sepsis, stroke, osteoporosis, osteoarthritis, ischemic reperfusion injury, cardiovascular disease, kidney disease, liver disease, ischemic injury, myocardial infarction, hypotension, hypertension, AIDS, myelodysplastic syndromes and other hematologic abnormalities, aplastic anemia, male pattern baldness, and bacterial, fungal, protozoan, and viral infections.

The presence of a CUB domain in both the murine and human TANGO 202 proteins indicates that this protein is involved in biological processes common to other CUB domain-containing proteins, such as developmental processes and binding to mediators. Therefore, TANGO 202 protein has a role in disorders which involve inappropriate developmental processes (e.g., abnormally high proliferation or un-differentiation of a differentiated tissue or abnormally low differentiation or proliferation of a non-developed or non-differentiated tissue) and modulation of cell growth, proliferation, survival, differentiation, and activity. Such disorders include, by way of example and not limitation, various cancers and birth and developmental defects.

Thus, proteins and nucleic acids of the invention which are identical to, similar to, or derived from human and murine TANGO 202 proteins and nucleic acids encoding them are useful for preventing, diagnosing, and treating, among others, vascular and cardiovascular disorders, hematological disorders, disorders related to mal-expression of growth factors, and cancer. Other uses for these proteins and nucleic acids of the invention relate to modulating cell growth (e.g., angiogenesis), proliferation (e.g., cancers), survival (e.g., apoptosis), differentiation (e.g., hematopoiesis), and activity (e.g., ligand-binding capacity). TANGO 202 proteins and nucleic acids encoding them are also useful for modulating cell dissociation and modulating migration of cells in extracellular matrices.

TANGO 234

A cDNA clone (designated jthsa104d11) encoding at least a portion of human TANGO 234 protein was isolated from a human fetal spleen cDNA library. The human TANGO 234 protein is predicted by structural analysis to be a transmembrane protein, although it can exist in a secreted form as well.

The full length of the cDNA encoding human TANGO 234 protein (FIG. 2; SEQ ID NO: 9) is 4628 nucleotide residues. The ORF of this cDNA, nucleotide residues 28 to 4386 of SEQ ID NO: 9 (i.e., SEQ ID NO: 10), encodes a 1453-amino acid transmembrane protein (FIG. 2; SEQ ID NO: 11).

The invention thus includes purified human TANGO 234 protein, both in the form of the immature 1453 amino acid residue protein (SEQ ID NO: 11) and in the form of the mature 1413 amino acid residue protein (SEQ ID NO: 13). Mature human TANGO 234 protein can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or it can be synthesized by generating immature TANGO 234 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature human TANGO 234 proteins, the invention includes fragments, derivatives, and variants of these TANGO 234 proteins, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 9 or some portion thereof, such as the portion which encodes mature TANGO 234 protein, immature TANGO 234 protein, or a domain of TANGO 234 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 234 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features, as indicated by the conservation of amino acid sequence between human TANGO 234 protein and bovine WC1 protein, as shown in FIGS. 2K through 2P, and the conservation of nucleotide sequence between the ORFs encoding human TANGO 234 protein and bovine WC1 protein, as shown in FIGS. 2Qi through 2Qxvii.

A common domain present in TANGO 234 proteins is a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 234 protein contains a signal sequence corresponding to amino acid residues 1 to 40 of SEQ ID NO: 11 (SEQ ID NO: 12). The signal sequence is cleaved during processing of the mature protein.

TANGO 234 proteins can include an extracellular domain. The human TANGO 234 protein extracellular domain is located from about amino acid residue 41 to about amino acid residue 1359 of SEQ ID NO: 3. TANGO 234 can alternately exist in a secreted form, such as a mature protein having the amino acid sequence of amino acid residues 41 to 1453 or residues 41 to about 1359 of SEQ ID NO: 11.

In addition, TANGO 234 include a transmembrane domain. In one embodiment, a TANGO 234 protein of the invention contains a transmembrane domain corresponding to about amino acid residues 1360 to 1383 of SEQ ID NO: 11 (SEQ ID NO: 15).

The present invention includes TANGO 234 proteins having a cytoplasmic domain, particularly including proteins having a carboxyl-terminal cytoplasmic domain. The human TANGO 234 cytoplasmic domain is located from about amino acid residue 1384 to amino acid residue 1453 of SEQ ID NO: 11 (SEQ ID NO: 16).

TANGO 234 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Table IV, as predicted by computerized sequence analysis of TANGO 234 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 234 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational modification sites listed in Table IV.

TABLE IV Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 11 Sequence N-glycosylation site 42 to 45 NGTD 78 to 81 NTTA 120 to 123 NESA 161 to 164 NNSC 334 to 337 NESF 377 to 380 NCSG 441 to 444 NESA 548 to 551 NESN 637 to 640 NAST 972 to 975 NESL 1013 to 1016 NVSD 1084 to 1087 NATV 1104 to 1107 NCTG 1161 to 1164 NGTW 1171 to 1174 NITT 1318 to 1321 NESF 1354 to 1357 NASS Glycosaminoglycan attachment site 558 to 561 SGWG 665 to 668 SGWG cAMP- or cGMP-dependent 1229 to 1232 RRIS protein kinase phosphorylation 1399 to 1402 RRGS site Protein kinase C phosphorylation 165 to 167 SGR site 268 to 270 TNR 379 to 381 SGR 419 to 421 SRR 469 to 471 SDK 506 to 508 STR 589 to 591 SNR 593 to 595 SGR 661 to 663 SCR 696 to 698 SSR 746 to 748 TER 805 to 807 SGR 815 to 817 TWR 959 to 961 SVR 1256 to 1258 SGR 1349 to 1351 SLK 1396 to 1398 STR Casein kinase II phosphorylation 44 to 47 TDLE site 71 to 74 TVCD 178 to 181 TICD 245 to 248 SHNE 253 to 256 TCYD 258 to 261 SDLE 319 to 322 SGSD 332 to 335 SGNE 392 to 395 TICD 439 to 442 TGNE 606 to 609 TVCD 622 to 625 SQLD 673 to 676 SHSE 686 to 689 SDME 760 to 763 TGGE 765 to 768 SLWD 818 to 821 SVCD 845 to 848 SVGD 857 to 860 TWAE 907 to 910 SQCD 923 to 926 SLCD 927 to 930 THWD 974 to 977 SLLD 1059 to 1062 TICD 1106 to 1109 TGTE 1145 to 1148 SETE 1233 to 1236 SPAE 1241 to 1244 TCED 1269 to 1272 TVCD 1402 to 1405 SLEE 1425 to 1428 TSDD N-myristoylation site 67 to 72 GQWGTV 90 to 95 GCPFSF 101 to 106 GQAVTR 119 to 124 GNESAL 133 to 138 GSHNCY 160 to 165 GNNSCS 197 to 202 GCPSSF 226 to 231 GNELAL 240 to 245 GNHDCS 267 to 272 GTNRCM 304 to 309 GCGTAL 328 to 333 GVSCSG 374 to 379 GSNNCS 411 to 416 GCPFSV 418 to 423 GSRRAK 440 to 445 GNESAL 465 to 470 GVICSD 547 to 552 GNESNI 588 to 593 GSNRCS 632 to 637 GMGLGN 668 to 673 GNNDCS 679 to 684 GVICSD 695 to 700 GSSRCA 712 to 717 GILCAN 720 to 725 GMNIAE 758 to 763 GCTGGE 853 to 858 GNGLTW 891 to 896 GVVCSR 944 to 949 GTALST 985 to 990 GAPPCI 992 to 997 GNTVSV 1078 to 1083 GCGVAF 1121 to 1126 GQHDCR 1132 to 1137 GVICSE 1162 to 1167 GTWGSV 1185 to 1190 GCGENG 1265 to 1270 GSWGTV 1288 to 1293 GCGSAL 1302 to 1307 GQGTGT 1331 to 1336 GQSDCG 1342 to 1347 GVRCSG 1422 to 1427 GTRTSD 1443 to 1438 GCEDAS 1444 to 1449 GVLPAS Amidation site 1167 to 1170 VGRR Speract receptor repeated (SRR) 53 to 90 See FIG. 2 domain signature 160 to 197 See FIG. 2 267 to 304 See FIG. 2 1041 to 1078 See FIG. 2 1251 to 1288 See FIG. 2 Scavenger receptor cysteine-rich  51 to 148 See FIG. 2 (SRCR) domain 158 to 255 See FIG. 2 265 to 362 See FIG. 2 372 to 469 See FIG. 2 479 to 576 See FIG. 2 586 to 683 See FIG. 2 693 to 790 See FIG. 2 798 to 895 See FIG. 2  903 to 1000 See FIG. 2 1039 to 1136 See FIG. 2 1146 to 1243 See FIG. 2 1249 to 1346 See FIG. 2

Among the domains that occur in TANGO 234 protein are SRR domains and SRCR domains. In one embodiment, the protein of the invention has at least one domain that is at least 55%, preferably at least about 65%, more preferably at least about 75%, yet more preferably at least about 85%, and most preferably at least about 95% identical to one of these domains. In other embodiments, the protein has at least two of the SRR and SRCR domains described herein in Table IV. In other embodiments, the protein has at least one SRR domain and at least one SRCR domain.

The SRR domain is named after a receptor domain identified in a sea urchin egg protein designated speract. The consensus sequence of this domain (using standard one-letter amino acid codes, wherein X is any amino acid residue) is as follows.

-G-X₅-G-X₂-E-X₆-W-G-X₂-C-X₃-(F or Y or W)-X₈-C-X₃-G-.

Speract is a transmembrane glycoprotein of 500 amino acid residues (Dangott et al. (1989) Proc. Natl. Acad. Sci. USA 86:2128-2132). Structurally, this receptor consists of a large extracellular domain of 450 residues, followed by a transmembrane region and a small cytoplasmic domain of 12 amino acid residues. The extracellular domain contains four repeats of an approximately 115 amino acid domain. There are 17 amino acid residues that are perfectly conserved in the four repeats in speract, including six cysteine residues, six glycine residues, and two glutamate residues. TANGO 234 has five SRR domains, in which 16 of the 17 conserved speract residues are present of four of the SRR domains and 15 are present in the remaining SRR domain. This domain is designated the speract receptor repeated domain. The amino acid sequence of mammalian macrophage scavenger receptor type I (MSRI) exhibits such a domain (Freeman et al. (1990) Proc. Natl. Acad. Sci. USA 87:8810-8814). MSRI proteins are membrane glycoproteins implicated in the pathologic deposition of cholesterol in arterial walls during atherogenesis. TANGO 234 is involved in one or more physiological processes related to cholesterol deposition and atherogenesis, as well as other vascular and cardiovascular disorders.

Scavenger receptor cysteine-rich (SRCR) domains are disulfide rich extracellular domains which are present in certain cell surface and secreted proteins. Proteins having SRCR domains exhibit diverse ligand binding specificity. For example, in addition to modified lipoproteins, some of these proteins bind a variety of surface components of pathogenic microorganisms, and some of the proteins bind apoptotic cells. SRCR domains are also involved in mediating immune development and response. Other SRCR-containing proteins are involved in binding of modified lipoproteins (e.g., oxidized low density lipoprotein {LDL}) by specialized macrophages, leading to the formation of macrophages filled with cholesteryl ester droplets (i.e., foam cells). TANGO 234 is involved in one or more physiological processes in which these other SRCR domain-containing proteins are involved, such as LDL uptake and metabolism, regulation of serum cholesterol level, atherogenesis, atherosclerosis, bacterial or viral infections, immune development, and generation and perseverance of immune responses.

WC1 is a ruminant protein having an SRCR domain. WC1 and gamma delta T-cell receptor are the only known gamma delta T-cell specific antigens. Antibodies which bind specifically with WC1 induce growth arrest in IL-2-dependent gamma delta T-cell and augment proliferation of gamma delta T-cells in an autologous mixed lymphocyte reaction or in the presence of anti-CD2 or anti-CD5 antibodies. Injection of antibodies which bind specifically with WC1 into calves results in long-lasting depletion of gamma delta T-cells. Furthermore, antibodies which bind specifically with WC1 can be used to purify gamma delta T-cells.

Gamma delta T-cells are involved in a variety of physiological processes. For example, these cells are potential mediators of allergic airway inflammation and lyme disease. Furthermore, these cells are involved in natural resistance to viral infections and can mediate autoimmune diseases. Elimination of gamma delta T-cells by injection of antibodies which bind specifically therewith can affect the outcomes of these disorders.

TANGO 234 is likely the human orthologue of ruminant protein WC1, and thus is involved with the physiological processes described above in humans. An alignment of the amino acid sequences of (human) TANGO 234 and bovine WC1 protein is shown in FIGS. 2K-2P. In this alignment (made using the ALIGN software {Myers and Miller (1989) CABIOS, ver. 2.0}; pam120.mat scoring matrix; gap penalties −12/−4), the proteins are 40.4% identical. An alignment of the nucleotide sequences of the ORFs encoding (human) TANGO 234 and bovine WC1 protein is shown in FIGS. 2Qi-2Qxvii. The two ORFs are 54.3% identical, as assessed using the same software and parameters.

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10: 1-6) predicted that human TANGO 234 protein includes a 40 amino acid signal peptide (amino acid residues 1 to 40 of SEQ ID NO: 11; SEQ ID NO: 12) preceding the mature TANGO 234 protein (amino acid residues 41 to 4386 of SEQ ID NO: 11; SEQ ID NO: 13). Human TANGO 234 protein includes an extracellular domain (amino acid residues 41 to 1359 of SEQ ID NO: 11; SEQ ID NO: 14); a transmembrane domain (amino acid residues 1360 to 1383 of SEQ ID NO: 11; SEQ ID NO: 15); and a cytoplasmic domain (amino acid residues 1384 to 1453 of SEQ ID NO: 11; SEQ ID NO: 16).

FIG. 2J depicts a hydrophilicity plot of human TANGO 234 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 40 of SEQ ID NO: 11 is the signal sequence of human TANGO 234 (SEQ ID NO: 12). The hydrophobic region which corresponds to amino acid residues 1360 to 1383 of SEQ ID NO: 11 is the transmembrane domain of human TANGO 234 (SEQ ID NO: 15). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 234 protein from about amino acid residue 225 to about amino acid residue 250 appears to be located at or near the surface of the protein, while the region from about amino acid residue 990 to about amino acid residue 1000 appears not to be located at or near the surface.

The predicted molecular weight of human TANGO 234 protein without modification and prior to cleavage of the signal sequence is about 159.3 kilodaltons. The predicted molecular weight of the mature human TANGO 234 protein without modification and after cleavage of the signal sequence is about 154.7 kilodaltons.

Chromosomal mapping to identify the location of the gene encoding human TANGO 234 protein indicated that the gene was located at chromosomal location h12p13 (with synteny to mo6). Flanking chromosomal markers include WI-6980 and GATA8A09.43. Nearby human loci include IBD2 (inflammatory bowel disease 2), FPF (familial periodic fever), and HPDR2 (hypophosphatemia vitamin D resistant rickets 2). Nearby genes are KLRC (killer cell receptor cluster), DRPLA (dentatorubro-pallidoluysian atrophy), GAPD (glyceraldehyde-3-phosphate)dehydrogenase, and PXR1 (peroxisome receptor 1). Murine chromosomal mapping indicated that the murine orthologue is located near the scr (scruffy) locus. Nearby mouse genes include drpla (dentatorubral phillidoluysian atrophy), prp (proline rich protein), and kap (kidney androgen regulated protein).

Northern analysis experiments indicated that mRNA corresponding to the cDNA encoding TANGO 234 is expressed in the tissues listed in Table V, wherein “++” indicates moderate expression, “+” indicates lower expression, and “−” indicates no detectable expression.

TABLE V Animal Tissue Relative Level of Expression Human spleen ++ fetal lung ++ lung + thymus + bone marrow − peripheral blood leukocytes −

Biological Function of TANGO 234 Proteins, Nucleic Acids, and Modulators Thereof

TANGO 234 proteins are involved in disorders which affect both tissues in which they are normally expressed and tissues in which they are normally not expressed. Based on the observation that TANGO 234 is expressed in human fetal lung, spleen, and, to a lesser extent in adult lung and thymus tissue, TANGO 234 protein is involved in one or more biological processes which occur in these tissues. In particular, TANGO 234 is involved in modulating growth, proliferation, survival, differentiation, and activity of cells including, but not limited to, lung, spleen, thymus bone marrow, hematopoietic, peripheral blood leukocytes, and fetal cells of the animal in which it is normally expressed. Thus, TANGO 234 has a role in disorders which affect these cells and their growth, proliferation, survival, differentiation, and activity. Expression of TANGO 234 in an animal is also involved in modulating growth, proliferation, survival, differentiation, and activity of cells and viruses which are foreign to the host (i.e., bacterial, fungal, and viral infections).

Homology of human TANGO 234 with bovine WC1 protein indicates that TANGO 234 has physiological functions in humans analogous to the functions of WC1 in ruminants. Thus, TANGO 234 is involved in modulating growth, proliferation, survival, differentiation, and activity of gamma delta T cells. For example, TANGO 234 affects the ability of gamma delta T cells to interact with chemokines such as interleukin-2. TANGO 234 therefore is involved in the physiological processes associated with allergic airway inflammation, lyme arthritis, resistance to viral infection, auto-immune diseases, and the like.

In addition, presence in TANGO 234 of SRR and SRCR domains indicates that TANGO 234 is involved in physiological functions identical or analogous to the functions performed by other proteins having such domains. For example, like other SRR domain-containing proteins, TANGO 234 modulates cholesterol deposition in arterial walls, and is thus involved in development and persistence of atherogenesis and arteriosclerosis, as well as other vascular and cardiovascular disorders. Like other SRCR domain-containing proteins, TANGO 234 is involved in uptake and metabolism of LDL, regulation of serum cholesterol level, and can modulate these processes as well as the processes of atherogenesis, arteriosclerosis, immune development, and generation and perseverance of immune responses to bacterial, fungal, and viral infections.

TANGO 265

A cDNA clone (designated jthsa079g01) encoding at least a portion of human TANGO 265 protein was isolated from a human fetal spleen cDNA library. The human TANGO 265 protein is predicted by structural analysis to be a transmembrane membrane protein, although it can exist in a secreted form as well.

The full length of the cDNA encoding human TANGO 265 protein (FIG. 3; SEQ ID NO: 17) is 3104 nucleotide residues. The ORF of this cDNA, nucleotide residues 32 to 2314 of SEQ ID NO: 17 (i.e., SEQ ID NO: 18), encodes a 761-amino acid transmembrane protein (FIG. 3; SEQ ID NO: 19).

The invention thus includes purified TANGO 265 protein, both in the form of the immature 761 amino acid residue protein (SEQ ID NO: 19) and in the form of the mature 730 amino acid residue protein (SEQ ID NO: 21). Mature TANGO 265 protein can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or it can be synthesized by generating immature TANGO 265 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature TANGO 265 proteins, the invention includes fragments, derivatives, and variants of TANGO 265 protein, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 17 or some portion thereof, such as the portion which encodes mature TANGO 265 protein, immature TANGO 265 protein, or a domain of TANGO 265 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 265 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features.

A common domain present in TANGO 265 proteins is a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 265 protein contains a signal sequence corresponding to amino acid residues 1 to 31 of SEQ ID NO: 19 (SEQ ID NO: 20). The signal sequence is cleaved during processing of the mature protein.

TANGO 265 proteins can also include an extracellular domain. The human TANGO 265 protein extracellular domain is located from about amino acid residue 32 to about amino acid residue 683 of SEQ ID NO: 17. TANGO 265 can alternately exist in a secreted form, such as a mature protein having the amino acid sequence of amino acid residues 32 to 761 or residues 32 to about 683 of SEQ ID NO: 19.

TANGO 265 proteins can also include a transmembrane domain. In one embodiment, a TANGO 265 protein of the invention contains a transmembrane domain corresponding to about amino acid residues 684 to 704 of SEQ ID NO: 19 (SEQ ID NO: 23).

In addition, TANGO 265 proteins include a cytoplasmic domain, particularly including proteins having a carboxyl-terminal cytoplasmic domain. The human TANGO 265 cytoplasmic domain is located from about amino acid residue 705 to amino acid residue 761 of SEQ ID NO: 19 (SEQ ID NO: 24).

TANGO 265 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Table VI, as predicted by computerized sequence analysis of TANGO 265 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 265 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational modification sites listed in Table VI.

TABLE IV Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 19 Sequence N-glycosylation site 120 to 123 NETQ 135 to 138 NVTH 496 to 499 NCSV 607 to 610 NGLS Glycosaminoglycan attachment site 70 to 73 SGDG cAMP- or cGMP-dependent 108 to 111 RKKS protein kinase phosphorylation 116 to 119 KKKS site 281 to 284 KKWT Protein kinase C phosphorylation 106 to 108 SDR site 262 to 264 TSR 361 to 363 TSR 366 to 368 TYR 385 to 387 SDK 533 to 535 SWK 555 to 557 SLR 721 to 723 TLR 738 to 740 SPK Casein kinase II phosphorylation 152 to 155 TFIE site 176 to 179 SPFD 250 to 253 TASE 342 to 345 SLLD 411 to 414 SGVE 498 to 501 SVYE 502 to 505 SCVD 574 to 577 SILE 738 to 741 SPKE 745 to 748 SASD N-myristoylation site 79 to 84 GAREAI 191 to 196 GMLYSG 331 to 336 GGTRSS 412 to 417 GVEYTR 437 to 442 GTTTGS 620 to 625 GLYQCW 671 to 676 GAALAA Sema domain  64 to 478 See FIG. 3

An exemplary domains which occurs in TANGO 265 proteins is a sema domain. In one embodiment, the protein of the invention has at least one domain that is at least 55%, preferably at least about 65%, more preferably at least about 75%, yet more preferably at least about 85%, and most preferably at least about 95% identical to one of the sema domains described herein in Table VI.

Sema domains occur in semaphorin proteins. Semaphorins are a large family of secreted and transmembrane proteins, some of which function as repellent signals during neural axon guidance. The sema domain and a variety of semaphorin proteins in which it occurs are described, for example, in Winberg et al. (1998 Cell 95:903-916). Sema domains also occur in human hepatocyte growth factor receptor (Swissprot Accession no. P08581) and the similar neuronal and epithelial transmembrane receptor protein (Swissprot Accession no. P51805). The presence of an sema domain in human TANGO 265 protein indicates that TANGO 265 is involved in one or more physiological processes in which the semaphorins are involved, has biological activity in common with one or more of the semaphorins, or both.

Human TANGO 265 protein exhibits considerable sequence similarity to murine semaphorin B protein (GenBank Accession no. X85991), as indicated herein in FIGS. 3F-3H. FIGS. 3F-3H depict an alignment of the amino acid sequences of human TANGO 265 protein (SEQ ID NO: 19) and murine semaphorin B protein (SEQ ID NO: 76). In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the amino acid sequences of the proteins are 82.3% identical. FIGS. 3I through 3T depict an alignment of the nucleotide sequences of cDNA encoding human TANGO 265 protein (SEQ ID NO: 17) and murine cDNA encoding semaphorin B protein (SEQ ID NO: 77). In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the nucleic acid sequences of the cDNAs are 76.2% identical. Thus, TANGO 265 is the human orthologue of murine semaphorin B and shares functional similarities to that protein.

It is known that semaphorins are bi-functional, capable of functioning either as attractive axonal guidance proteins or as repellent axonal guidance proteins (Wong et al. (1997) Development 124:3597-3607). Furthermore, semaphorins bind with neuronal cell surface proteins designated plexins, which are expressed on both neuronal cells and cells of the immune system (Comeau et al. (1998) Immunity 8:473-482; Jin and Strittmatter (1997) J. Neurosci. 17:6256-6263).

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that human TANGO 265 protein includes a 31 amino acid signal peptide (amino acid residues 1 to 31 of SEQ ID NO: 19; SEQ ID NO: 20) preceding the mature TANGO 265 protein (amino acid residues 32 to 761 of SEQ ID NO: 19; SEQ ID NO: 21). Human TANGO 265 protein includes an extracellular domain (amino acid residues 32 to 683 of SEQ ID NO: 19; SEQ ID NO: 22); a transmembrane domain (amino acid residues 684 to 704 of SEQ ID NO: 19; SEQ ID NO: 23); and a cytoplasmic domain (amino acid residues 705 to 761 of SEQ ID NO: 19; SEQ ID NO: 24).

FIG. 3U depicts a hydrophilicity plot of human TANGO 265 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 31 of SEQ ID NO: 19 is the signal sequence of human TANGO 265 (SEQ ID NO: 20). The hydrophobic region which corresponds to amino acid residues 684 to 704 of SEQ ID NO: 19 is the transmembrane domain of human TANGO 265 (SEQ ID NO: 23). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 265 protein from about amino acid residue 350 to about amino acid residue 375 appears to be located at or near the surface of the protein, while the region from about amino acid residue 230 to about amino acid residue 250 appears not to be located at or near the surface.

The predicted molecular weight of human TANGO 265 protein without modification and prior to cleavage of the signal sequence is about 83.6 kilodaltons. The predicted molecular weight of the mature human TANGO 265 protein without modification and after cleavage of the signal sequence is about 80.2 kilodaltons.

Chromosomal mapping was performed by computerized comparison of TANGO 265 cDNA sequences against a chromosomal mapping database in order to identify the approximate location of the gene encoding human TANGO 265 protein. This analysis indicated that the gene was located on chromosome 1 between markers D1S305 and D1S2635.

Biological Function of TANGO 265 Proteins, Nucleic Acids, and Modulators Thereof

TANGO 265 proteins are involved in disorders which affect both tissues in which they are normally expressed and tissues in which they are normally not expressed. Based on the observation that TANGO 265 is expressed in human fetal spleen, involvement of TANGO 202 protein in immune system development and modulation is indicated.

The presence of the sema domain in TANGO 265 indicates that this protein is involved in development of neuronal and epithelial tissues and also functions as a repellant protein which guides axonal development. TANGO 265 modulates nerve growth and regeneration and also modulates growth and regeneration of other epithelial tissues.

The observation that TANGO 265 shares significant identity with murine semaphorin B suggests that it has activity identical or analogous to the activity of this protein. These observations indicate that TANGO 265 modulates growth, proliferation, survival, differentiation, and activity of neuronal cells and immune system cells. Thus, TANGO 265 protein is useful, for example, for guiding neural axon development, for modulating differentiation of cells of the immune system, for modulating cytokine production by cells of the immune system, for modulating reactivity of cells of the immune system toward cytokines, for modulating initiation and persistence of an inflammatory response, and for modulating proliferation of epithelial cells.

TANGO 273

A cDNA clone (designated jthoc028g06) encoding at least a portion of human TANGO 273 protein was isolated from a lipopolysaccharide-(LPS-)stimulated human osteoblast cDNA library. The corresponding murine cDNA clone (designated jtmoa001c04) was isolated from an LPS-stimulated murine osteoblast cDNA library. The human and murine TANGO 273 proteins are predicted by structural analysis to be transmembrane proteins.

The full length of the cDNA encoding human TANGO 273 protein (FIG. 4; SEQ ID NO: 25) is 2964 nucleotide residues. The ORF of this cDNA, nucleotide residues 135 to 650 of SEQ ID NO: 25 (i.e., SEQ ID NO: 26), encodes a 172-amino acid transmembrane protein (FIG. 4; SEQ ID NO: 27).

The invention thus includes purified human TANGO 273 protein, both in the form of the immature 172 amino acid residue protein (SEQ ID NO: 27) and in the form of the mature 150 amino acid residue protein (SEQ ID NO: 29). The invention also includes purified murine TANGO 273 protein, both in the form of the immature 172 amino acid residue protein (SEQ ID NO: 74) and in the form of the mature 150 amino acid residue protein (SEQ ID NO: 44). Mature human or murine TANGO 273 proteins can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or they can be synthesized by generating immature TANGO 273 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature human and murine TANGO 273 proteins, the invention includes fragments, derivatives, and variants of these TANGO 273 proteins, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 25 or some portion thereof or SEQ ID NO: 73 or some portion thereof, such as the portion which encodes mature TANGO 273 protein, immature TANGO 273 protein, or a domain of TANGO 273 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 273 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features. This family includes, by way of example, the human and murine TANGO 273 proteins.

A common domain of TANGO 273 proteins is a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 273 protein contains a signal sequence corresponding to amino acid residues 1 to 22 of SEQ ID NO: 27 (SEQ ID NO: 28) or to amino acid residues 1 to 22 of SEQ ID NO: 74. The signal sequence is cleaved during processing of the mature protein.

TANGO 273 proteins can also include an extracellular domain. The human TANGO 273 protein extracellular domain is located from about amino acid residue 23 to about amino acid residue 60 of SEQ ID NO: 27, and the murine TANGO 273 protein extracellular domain is located from about amino acid residue 23 to about amino acid residue 60 of SEQ ID NO: 74.

The present invention also includes TANGO 273 proteins having a transmembrane domain. As used herein, a “transmembrane domain” refers to an amino acid sequence having at least about 15 to 30 amino acid residues in length and which contains at least about 65-70% hydrophobic amino acid residues such as alanine, leucine, phenylalanine, protein, tyrosine, tryptophan, or valine. In a preferred embodiment, a transmembrane domain contains at least about 15 to 20 amino acid residues, preferably about 20 to 25 amino acid residues, and has at least about 60-80%, more preferably 65-75%, and more preferably at least about 70% hydrophobic residues. Thus, in one embodiment, a human TANGO 273 protein of the invention contains a transmembrane domain corresponding to about amino acid residues 61 to 81 of SEQ ID NO: 27 (SEQ ID NO: 31). In another embodiment, a murine TANGO 273 protein of the invention contains a transmembrane domain corresponding to about amino acid residues 61 to 81 of SEQ ID NO: 74.

In addition, TANGO 273 proteins include a cytoplasmic domain. The human TANGO 273 cytoplasmic domain is located from about amino acid residue 82 to amino acid residue 172 of SEQ ID NO: 27 (SEQ ID NO: 32), and the murine TANGO 273 cytoplasmic domain is located from about amino acid residue 82 to amino acid residue 172 of SEQ ID NO: 74.

TANGO 273 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Tables VII and VIII, as predicted by computerized sequence analysis of human and murine TANGO 273 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 273 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2, 3, 4, 5, or all 6 of the post-translational modification sites listed in Table VII. In other embodiments, the protein of the invention has at least 1, 2, 3, 4, 5, 6, or all 7 of the post-translational modification sites listed in Table VIII.

TABLE VII Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 27 Sequence N-glycosylation site  97 to 100 NVSY Casein kinase II phosphorylation 41 to 44 SYED site N-myristoylation site 31 to 36 GLYPTY 47 to 52 GSRCCV 70 to 75 GVLFCC 131 to 136 GNSMAM Src Homology 3 (SR3) 86 to 90 YPPPL domain binding site 103 to 107 QPPNP 113 to 117 QPGPP 121 to 125 DPGGP 140 to 145 VPPNSP 151 to 155 CPPPP 160 to 164 TPPPP

TABLE VIII Type of Potential Modification Site Amino Acid Residues Amino Acid or Domain of SEQ ID NO: 74 Sequence N-glycosylation site  97 to 100 NVSY Casein kinase II phosphorylation 41 to 44 SYED site N-myristoylation site 31 to 36 GLYPTY 47 to 52 GSRCCV 70 to 75 GVLFCC 131 to 136 GNTMAM Src Homology 3 (SH3) 86 to 90 YPPPL domain binding site 103 to 107 QPPNP 115 to 119 GPPYY 121 to 125 DPGGP 141 to 145 QPNSP 151 to 155 YPPPP 160 to 164 TPPPP Amidation site 1 to 4 MGRR

The amino acid sequence of TANGO 273 protein includes about seven potential proline-rich Src homology 3 (SH3) domain binding sites nearer the cytoplasmic portion of the protein. SH3 domains mediate specific assembly of protein complexes, presumably by interacting with proline-rich protein domains (Morton and Campbell (1994) Curr. Biol. 4:615-617). SH3 domains also mediate interactions between proteins involved in transmembrane signal transduction. Coupling of proteins mediated by SH3 domains has been implicated in a variety of physiological systems, including those involving regulation of cell growth and proliferation, endocytosis, and activation of respiratory burst.

SH3 domains have been described in the art (e.g., Mayer et al. (1988) Nature 332:272-275; Musacchio et al. (1992) FEBS Lett. 307:55-61; Pawson and Schlessinger (1993) Curr. Biol. 3:434-442; Mayer and Baltimore (1993) Trends Cell Biol. 3:8-13; Pawson (1993) Nature 373:573-580), and occur in a variety of cytoplasmic proteins, including several (e.g., protein tyrosine kinases) involved in transmembrane signal transduction. Among the proteins in which one or more SH3 domains occur are protein tyrosine kinases such as those of the Src, Abl, Bkt, Csk and ZAP70 families, mammalian phosphatidylinositol-specific phospholipases C-gamma-1 and -2, mammalian phosphatidylinositol 3-kinase regulatory p85 subunit, mammalian Ras GTPase-activating protein (GAP), proteins which mediate binding of guanine nucleotide exchange factors and growth factor receptors (e.g., vertebrate GRB2, Caenorhabditis elegans sem-5, and Drosophila DRK proteins), mammalian Vav oncoprotein, guanidine nucleotide releasing factors of the CDC 25 family (e.g., yeast CDC25, yeast SCD25, and fission yeast ste6 proteins), MAGUK proteins (e.g., mammalian light junction protein ZO-1, vertebrate erythrocyte membrane protein p55, C. elegans protein lin-2, rat protein CASK, and mammalian synaptic proteins SAP90/PSD-95, CHAPSYN-110/PSD-93, SAP97/DLG1, and SAP102), proteins which interact with vertebrate receptor protein tyrosine kinases (e.g., mammalian cytoplasmic protein Nck and oncoprotein Crk), chicken Src substrate p80/85 protein (cortactin), human hemopoietic lineage cell specific protein Hs1, mammalian dihydrouridine-sensitive L-type calcium channel beta subunit, human myasthenic syndrome antigen B (MSYB), mammalian neutrophil cytosolic activators of NADPH oxidase (e.g., p47 {NCF-1}, p67 {NCF-2}, and C. elegans protein B0303.7) myosin heavy chains (MYO3) from amoebae, from slime molds, and from yeast, vertebrate and Drosophila spectrin and fodrin alpha chain proteins, human amphiphysin, yeast actin-binding proteins ABP1 and SLA3, yeast protein BEM1, fission yeast protein scd2 (ral3), yeast BEM1-binding proteins BOI2 (BEB1) and BOB1 (BOI1), yeast fusion protein FUS1, yeast protein RSV167, yeast protein SSU81, yeast hypothetical proteins YAR014c, YFR024c, YHL002w, YHR016c, YJL020C, and YHR114w, hypothetical fission yeast protein SpAC12C2.05c, and C. elegans hypothetical protein F42H10.3. Of these proteins, multiple SH3 domains occur in vertebrate GRB2 protein, C. elegans sem-5 protein, Drosophila DRK protein, oncoprotein Crk, mammalian neutrophil cytosolic activators of NADPH oxidase p47 and p67, yeast protein BEM1, fission yeast protein scd2, yeast hypothetical protein YHR114w, mammalian cytoplasmic protein Nck, C. elegans neutrophil cytosolic activator of NADPH oxidase B0303.7, and yeast actin-binding protein SLA1. Of these proteins, three or more SH3 domains occur in mammalian cytoplasmic protein Nck, C. elegans neutrophil cytosolic activator of NADPH oxidase B0303.7, and yeast actin-binding protein SLA1. The presence of SH3 domain binding sites in TANGO 273 indicates that TANGO 273 interacts with one or more of these and other SH3 domain-containing proteins and is thus involved in physiological processes in which one or more of these or other SH3 domain-containing proteins are involved.

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that human TANGO 273 protein includes a 22 amino acid signal peptide (amino acid residues 1 to 22 of SEQ ID NO: 27; SEQ ID NO: 28) preceding the mature TANGO 273 protein (amino acid residues 23 to 172 of SEQ ID NO: 27; SEQ ID NO: 29). Human TANGO 273 protein includes an extracellular domain (amino acid residues 23 to 60 of SEQ ID NO: 27; SEQ ID NO: 30); a transmembrane domain (amino acid residues 61 to 81 of SEQ ID NO: 27; SEQ ID NO: 31); and a cytoplasmic domain (amino acid residues 82 to 172 of SEQ ID NO: 27; SEQ ID NO: 32).

FIG. 4I depicts a hydrophilicity plot of human TANGO 273 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 22 of SEQ ID NO: 27 is the signal sequence of human TANGO 273 (SEQ ID NO: 28). The hydrophobic region which corresponds to amino acid residues 61 to 81 of SEQ ID NO: 27 is the transmembrane domain of human TANGO 273 (SEQ ID NO: 31). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 273 protein from about amino acid residue 100 to about amino acid residue 120 appears to be located at or near the surface of the protein, while the region from about amino acid residue 130 to about amino acid residue 140 appears not to be located at or near the surface.

Chromosomal mapping was performed by computerized comparison of TANGO 273 cDNA sequences against a chromosomal mapping database in order to identify the approximate location of the gene encoding human TANGO 273 protein. This analysis indicated that the gene was located on chromosome 7 between markers D7S2467 and D7S2552.

The predicted molecular weight of human TANGO 273 protein without modification and prior to cleavage of the signal sequence is about 19.2 kilodaltons. The predicted molecular weight of the mature human TANGO 273 protein without modification and after cleavage of the signal sequence is about 16.8 kilodaltons.

Northern analysis experiments indicated that mRNA corresponding to the cDNA encoding TANGO 273 is expressed in the tissues listed in Table VIIa, wherein “++” indicates moderate expression and “+” indicates lower expression.

TABLE VIIa Animal Tissue Relative Level of Expression Human heart ++ brain ++ skeletal muscle ++ pancreas ++ placenta + lung + liver + kidney +

The full length of the cDNA encoding murine TANGO 273 protein (FIG. 4; SEQ ID NO: 72) is 2915 nucleotide residues. The ORF of this cDNA, nucleotide residues 137 to 650 of SEQ ID NO: 72 (i.e., SEQ ID NO: 73), encodes a 172-amino acid transmembrane protein (FIG. 4; SEQ ID NO: 74).

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that murine TANGO 273 protein includes a 22 amino acid signal peptide (amino acid residues 1 to 22 of SEQ ID NO: 74) preceding the mature TANGO 273 protein (amino acid residues 23 to 172 of SEQ ID NO: 74; SEQ ID NO: 44). Murine TANGO 273 protein includes an extracellular domain (amino acid residues 23 to 60 of SEQ ID NO: 74); a transmembrane domain (amino acid residues 61 to 81 of SEQ ID NO: 74); and a cytoplasmic domain (amino acid residues 82 to 172 of SEQ ID NO: 74).

FIG. 4J depicts a hydrophilicity plot of murine TANGO 273 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 22 of SEQ ID NO: 74 is the signal sequence of murine TANGO 273. As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of murine TANGO 273 protein from about amino acid residue 100 to about amino acid residue 120 appears to be located at or near the surface of the protein, while the region from about amino acid residue 130 to about amino acid residue 140 appears not to be located at or near the surface.

The predicted molecular weight of murine TANGO 273 protein without modification and prior to cleavage of the signal sequence is about 19.4 kilodaltons. The predicted molecular weight of the mature murine TANGO 273 protein without modification and after cleavage of the signal sequence is about 17.1 kilodaltons.

In situ analysis of murine TANGO 273 mRNA indicated that TANGO 273 is expressed with central nervous system (CNS) tissues during embryogenesis and into adulthood. Expression of TANGO 273 is widely observed in murine CNS tissues, including brain, spinal cord, eye, and olfactory epithelium at all embryonic ages examined (i.e., at embryonic days 13.5, 14.5, 15.5, 16.5, and 18.5 and at post-natal day 1.5).

Human and murine TANGO 273 cDNA sequences exhibit significant nucleotide sequence identity with an expressed sequence tag (EST) isolated from a library of ESTs corresponding to proteins secreted from prostate tissue, as described in PCT publication number WO 99/06550, published Feb. 11, 1999.

Human and murine TANGO 273 proteins exhibit considerable sequence similarity, as indicated herein in FIG. 4H. FIG. 4H depicts an alignment of human and murine TANGO 273 protein amino acid sequences (SEQ ID NOs: 27 and 74, respectively). In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the proteins are 89.5% identical. Alignment of the ORF encoding human TANGO 273 protein and the ORF encoding murine TANGO 273 protein using the same software and parameters indicated that the nucleotide sequences are 84.1% identical.

Biological Function of TANGO 273 Proteins, Nucleic Acids, and Modulators Thereof

cDNAs encoding the human and murine TANGO 273 proteins were each isolated from LPS-stimulated osteoblast cDNA libraries. These proteins are involved in bone-related metabolism, homeostasis, and development disorders. Thus, proteins and nucleic acids of the invention which are identical to, similar to, or derived from human and murine TANGO 273 proteins and nucleic acids encoding them are useful for preventing, diagnosing, and treating, among others, bone-related disorders such as osteoporosis, cancer, skeletal development disorders, bone fragility, and the like.

Expression of TANGO 273 in heart, brain, skeletal muscle, and pancreas, placenta, lung, liver, and kidney tissues is an indication that TANGO 273 proteins, nucleic acids encoding them, and agents that modulate activity or expression of either of these can be used to modulate growth, proliferation, survival, differentiation, adhesion, and activity of cells of these tissues, or to prognosticate, diagnose, and treat one or more disorders which affect these tissues.

The fact that TANGO 273 is expressed at high levels in neurological tissues is an indication that TANGO 273 proteins, nucleic acids, and modulators thereof can be used to modulate proliferation, differentiation, or function of neurological cells in these tissues (e.g., neuronal cells). Thus, TANGO 273 proteins, nucleic acids, and modulators thereof can be used to prognosticate, diagnose, and treat one or more neurological disorders. Examples of such disorders include CNS disorders, CNS-related disorders, focal brain disorders, global-diffuse cerebral disorders, and other neurological and cerebrovascular disorders.

CNS disorders include, but are not limited to cognitive and neurodegenerative disorders such as Alzheimer's disease, senile dementia, Huntington's disease, amyotrophic lateral sclerosis, and Parkinson's disease, as well as Gilles de la Tourette's syndrome, autonomic function disorders such as hypertension and sleep disorders (e.g., insomnia, hypersomnia, parasomnia, and sleep apnea); neuropsychiatric disorders (e.g., schizophrenia, schizoaffective disorder, attention deficit disorder, dysthymic disorder, major depressive disorder, mania, and obsessive-compulsive disorder); psychoactive substance use disorders; anxiety; panic disorder; and bipolar affective disorders (e.g., severe bipolar affective disorder and bipolar affective disorder with hypomania and major depression).

CNS-related disorders include disorders associated with developmental, cognitive, and autonomic neural and neurological processes, such as pain, appetite, long term memory, and short term memory.

Exemplary focal brain disorders include aphasia, apraxia, agnosia, and amnesias (e.g., posttraumatic amnesia, transient global amnesia, and psychogenic amnesia). Global-diffuse cerebral disorders with which TANGO 273 can be associated include coma, stupor, obtundation, and disorders of the reticular formation.

Other neurological disorders with which TANGO 273 can be associated include ischemic syndromes (e.g., stroke), hypertensive encephalopathy, hemorrhagic disorders, and disorders involving aberrant function of the blood-brain barrier (e.g., CNS infections such as meningitis and encephalitis, aseptic meningitis, metastasis of non-CNS tumor cells into the CNS, various pain disorders such as migraine, blindness and other vision problems, and CNS-related adverse drug reactions such as head pain, sleepiness, and confusion). TANGO 273 proteins, nucleic acids encoding them, and agents that modulate activity or expression of either of these can be used to prognosticate, diagnose, and treat one or more of these disorders.

Developmental regulation of TANGO 273 expression in fetal neurological tissues, as described herein, is an indication that TANGO 273 proteins, nucleic acids, and modulators thereof can be used to prognosticate, diagnose, and treat one or more disorders which involve aberrant fetal neurological development. Examples of such disorders include blindness, deafness, fetal death, mental retardation, dysraphia, anencephaly, malformation of cerebral hemispheres, encephalocele, porencephaly, hydranencephaly, hydrocephalus, and spina bifida.

The fact that TANGO 273 is expressed in tissues which were exposed to LPS indicates that TANGO 273 mediates one or more physiological responses of cells to bacterial infection. Thus, TANGO 273 is involved in one or more of detection of bacteria in a tissue in which it is expressed, movement of cells with relation to sites of bacterial infection, production of biological molecules which inhibit bacterial infection, and production of biological molecules which alleviate cellular or other physiological damage wrought by bacterial infection.

Presence in TANGO 273 protein of multiple SH3 domain binding sites indicates that TANGO 273 protein interacts with one or more SH3 domain-containing proteins. Thus, TANGO 273 protein mediates binding of proteins (i.e., binding of proteins to TANGO 273 and to one another to form protein complexes) in cells in which it is expressed. TANGO 273 is also involved in transduction of signals between the exterior environment of cells (i.e., including from other cells) and the interior of cells in which it is expressed. TANGO 273 mediates regulation of cell growth and proliferation, endocytosis, activation of respiratory burst, and other physiological processes triggered by transmission of a signal via a protein with which TANGO 273 interacts.

Sequence similarity of TANGO 273 cDNA with an EST expressed in prostate tissue indicates that TANGO 273 can be expressed in prostate tissue, and can thus be involved in disorders of the prostate. Thus, TANGO 273 proteins, nucleic acids encoding them, and agents that modulate activity or expression of either of these can be used to treat prostate disorders. Examples of prostate disorders which can be treated in this manner include inflammatory prostatic diseases (e.g., acute and chronic prostatitis and granulomatous prostatitis), prostatic hyperplasia (e.g., benign prostatic hypertrophy or hyperplasia), and prostate tumors (e.g., carcinomas).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat cardiovascular disorders, such as ischemic heart disease (e.g., angina pectoris, myocardial infarction, and chronic ischemic heart disease), hypertensive heart disease, pulmonary heart disease, valvular heart disease (e.g., rheumatic fever and rheumatic heart disease, endocarditis, mitral valve prolapse, and aortic valve stenosis), congenital heart disease (e.g., valvular and vascular obstructive lesions, atrial or ventricular septal defect, and patent ductus arteriosus), or myocardial disease (e.g., myocarditis, congestive cardiomyopathy, and hypertrophic cardiomyopathy).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat disorders of the brain, such as cerebral edema, hydrocephalus, brain herniations, iatrogenic disease (due to, e.g., infection, toxins, or drugs), inflammations (e.g., bacterial and viral meningitis, encephalitis, and cerebral toxoplasmosis), cerebrovascular diseases (e.g., hypoxia, ischemia, and infarction, intracranial hemorrhage and vascular malformations, and hypertensive encephalopathy), and tumors (e.g., neuroglial tumors, neuronal tumors, tumors of pineal cells, meningeal tumors, primary and secondary lymphomas, intracranial tumors, and medulloblastoma), and to treat injury or trauma to the brain.

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat disorders of skeletal muscle, such as muscular dystrophy (e.g., Duchenne muscular dystrophy, Becker muscular dystrophy, Emery-Dreifuss muscular dystrophy, limbgirdle muscular dystrophy, facioscapulohumeral muscular dystrophy, myotonic dystrophy, oculopharyngeal muscular dystrophy, distal muscular dystrophy, and congenital muscular dystrophy), motor neuron diseases (e.g., amyotrophic lateral sclerosis, infantile progressive spinal muscular atrophy, intermediate spinal muscular atrophy, spinal bulbar muscular atrophy, and adult spinal muscular atrophy), myopathies (e.g., inflammatory myopathies such as dermatomyositis and polymyositis, myotonia congenita, paramyotonia congenita, central core disease, nemaline myopathy, myotubular myopathy, and periodic paralysis), and metabolic diseases of muscle (e.g., phosphorylase deficiency, acid maltase deficiency, phosphofructokinase deficiency, debrancher enzyme deficiency, mitochondrial myopathy, carnitine deficiency, carnitine palmityl transferase deficiency, phosphoglycerate kinase deficiency, phosphoglycerate mutase deficiency, lactate dehydrogenase deficiency, and myoadenylatc deaminase deficiency).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat pancreatic disorders, such as pancreatitis (e.g., acute hemorrhagic pancreatitis and chronic pancreatitis), pancreatic cysts (e.g., congenital cysts, pseudocysts, and benign or malignant neoplastic cysts), pancreatic tumors (e.g., pancreatic carcinoma and adenoma), diabetes mellitus (e.g., insulin- and non-insulin-dependent types, impaired glucose tolerance, and gestational diabetes), or islet cell tumors (e.g., insulinomas, adenomas, Zollinger-Ellison syndrome, glucagonomas, and somatostatinoma).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat placental disorders, such as toxemia of pregnancy (e.g., preeclampsia and eclampsia), placentitis, or spontaneous abortion.

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat pulmonary disorders, such as atelectasis, cystic fibrosis, rheumatoid lung disease, pulmonary congestion or edema, chronic obstructive airway disease (e.g., emphysema, chronic bronchitis, bronchial asthma, and bronchiectasis), diffuse interstitial diseases (e.g., sarcoidosis, pneumoconiosis, hypersensitivity pneumonitis, Goodpasture's syndrome, idiopathic pulmonary hemosiderosis, pulmonary alveolar proteinosis, desquamative interstitial pneumonitis, chronic interstitial pneumonia, fibrosing alveolitis, hamman-rich syndrome, pulmonary eosinophilia, diffuse interstitial fibrosis, Wegener's granulomatosis, lymphomatoid granulomatosis, and lipid pneumonia), or tumors (e.g., bronchogenic carcinoma, bronchioalveolar carcinoma, bronchial carcinoid, hamartoma, and mesenchymal tumors).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat hepatic (liver) disorders, such as jaundice, hepatic failure, hereditary hyperbilirubinemias (e.g., Gilbert's syndrome, Crigler-Naijar syndromes, and Dubin-Johnson and Rotor's syndromes), hepatic circulatory disorders (e.g., hepatic vein thrombosis and portal vein obstruction and thrombosis) hepatitis (e.g., chronic active hepatitis, acute viral hepatitis, and toxic and drug-induced hepatitis) cirrhosis (e.g., alcoholic cirrhosis, biliary cirrhosis, and hemochromatosis), or malignant tumors (e.g., primary carcinoma, hepatoblastoma, and angiosarcoma).

In another example, TANGO 273 polypeptides, nucleic acids, or modulators thereof, can be used to treat renal (kidney) disorders, such as glomerular diseases (e.g., acute and chronic glomerulonephritis, rapidly progressive glomerulonephritis, nephrotic syndrome, focal proliferative glomerulonephritis, glomerular lesions associated with systemic disease such as systemic lupus erythematosus, Goodpasture's syndrome, multiple myeloma, diabetes, neoplasia, sickle cell disease, and chronic inflammatory diseases), tubular diseases (e.g., acute tubular necrosis and acute renal failure, polycystic renal disease, medullary sponge kidney, medullary cystic disease, nephrogenic diabetes, and renal tubular acidosis), tubulointerstitial diseases (e.g., pyelonephritis, drug and toxin induced tubulointerstitial nephritis, hypercalcemic nephropathy, and hypokalemic nephropathy) acute and rapidly progressive renal failure, chronic renal failure, nephrolithiasis, vascular diseases (e.g., hypertension and nephrosclerosis, microangiopathic hemolytic anemia, atheroembolic renal disease, diffuse cortical necrosis, and renal infarcts), or tumors (e.g., renal cell carcinoma and nephroblastoma).

TANGO 286

A cDNA clone (designated jthkf042e03) encoding at least a portion of human TANGO 286 protein was isolated from a human keratinocyte cDNA library. The human TANGO 286 protein is predicted by structural analysis to be a secreted protein.

The full length of the cDNA encoding TANGO 286 protein (FIG. 5; SEQ ID NO: 33) is 1980 nucleotide residues. The ORF of this cDNA, nucleotide residues 133 to 1497 of SEQ ID NO: 33 (i.e., SEQ ID NO: 34), encodes a 455-amino acid secreted protein (FIG. 5; SEQ ID NO: 35).

The invention thus includes purified TANGO 286 protein, both in the form of the immature 455 amino acid residue protein (SEQ ID NO: 35) and in the form of the mature 432 amino acid residue protein (SEQ ID NO: 37). Mature TANGO 286 protein can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or it can be synthesized by generating immature TANGO 286 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature TANGO 286 proteins, the invention includes fragments, derivatives, and variants of these TANGO 286 proteins, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 33 or some portion thereof, such as the portion which encodes mature TANGO 286 protein, immature TANGO 286 protein, or a domain of TANGO 286 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 286 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features.

A common domain of TANGO 286 proteins is a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 286 protein contains a signal sequence corresponding to amino acid residues 1 to 23 of SEQ ID NO: 35 (SEQ ID NO: 36). The signal sequence is cleaved during processing of the mature protein.

TANGO 286 is a secreted soluble protein (i.e., a secreted protein having a single extracellular domain), as indicated by computerized sequence analysis and comparison of the amino acid sequence of TANGO 286 with related proteins, such as the soluble proteins designated bactericidal permeability increasing (BPI) protein and recombinant endotoxin neutralizing polypeptide (RENP).

TANGO 286 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Table IX, as predicted by computerized sequence analysis of TANGO 286 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 286 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational modification sites listed in Table IX.

TABLE IX Type of Potential Modification Amino Acid Residues Amino Acid Site or Domain of SEQ ID NO: 35 Sequence N-glycosylation site 79 to 82 NFSN 92 to 95 NTSL 113 to 116 NIST 161 to 164 NLST 173 to 176 NYTL 205 to 208 NLTD 249 to 252 NLTL 303 to 306 NFTL 320 to 323 NSTV 363 to 366 NRSN Protein kinase C phosphorylation 35 to 37 TQR site 362 to 364 SNR 429 to 431 SSK Casein kinase II phosphorylation 63 to 66 SGSE site 130 to 133 SFAE 163 to 166 STLE 169 to 172 TKID 175 to 178 TLLD 183 to 186 SSPE 253 to 256 STEE 321 to 324 STVE 365 to 368 SNIE 409 to 412 SDIE N-myristoylation site 42 to 47 GYQAGM 269 to 274 GNVLSR Lipid-binding serum glycoprotein  12 to 427 see FIG. 5 domain

Certain lipid-binding serum glycoproteins, such as LPS-binding protein (LBP), bactericidal permeability-increasing protein (BPI), cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP), share regions of sequence similarity which are herein designated a lipid-binding serum glycoprotein domain (Schumann et al., (1990) Science 249:1429-1431; Gray et al., (1989) J. Biol. Chem. 264:9505-9509; Day et al., (1994) J. Biol. Chem. 269:9388-9391). The consensus pattern of lipid-binding serum glycoprotein domains is as follows (using standard single letter amino acid abbreviations wherein X is any amino acid residue).

-(P or A)-(G or A)-(L or I or V or M or C)-X₂-R-(I or V)-(S or T)-X₃-L-X_((4 or 5))-(E or Q)-X₄-(L or I or V or M)-X_((0 or 1))-(E or Q or K)-X₈-P-

(e.g., amino acid residues 28-60 of SEQ ID NO: 35).

Proteins in which a lipid-binding serum glycoprotein domain occurs are often structurally related and exhibit related physiological activities. LBP binds to lipid A moieties of bacterial LPS and, once bound thereto, induces secretion of α-tumor necrosis factor, apparently by interacting with the CD14 receptor. BPI also binds LPS and exerts a cytotoxic effect on Gram-negative bacteria (Elsbach, (1998) J. Leukoc. Biol. 64:14-18). CETP is involved in transfer of insoluble cholesteryl esters during reverse cholesterol transport. PLTP appears to be involved in phospholipid transport and modulation of serum HDL particles.

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that TANGO 286 protein includes a 23 amino acid signal peptide (amino acid residues 1 to 23 of SEQ ID NO: 35; SEQ ID NO: 36) preceding the mature TANGO 286 protein (amino acid residues 24 to 455 of SEQ ID NO: 35; SEQ ID NO: 37). Human TANGO 286 protein is a secreted soluble protein.

FIG. 5E depicts a hydrophilicity plot of TANGO 286 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 286 protein from about amino acid residue 420 to about amino acid residue 435 appears to be located at or near the surface of the protein, while the region from about amino acid residue 325 to about amino acid residue 345 appears not to be located at or near the surface.

The predicted molecular weight of TANGO 286 protein without modification and prior to cleavage of the signal sequence is about 50.9 kilodaltons. The predicted molecular weight of the mature TANGO 286 protein without modification and after cleavage of the signal sequence is about 48.2 kilodaltons.

The gene encoding human TANGO 286 protein was determined to be located on chromosome 22 by comparison of matching genomic clones such as the clones assigned GenBank Accession numbers W16806 and AL021937.

A portion of TANGO 286 protein exhibits significant amino acid homology with a region of the human chromosome region 22q12-13 genomic nucleotide sequence having GenBank Accession number AL021937. Alignment of a 45 kilobase nucleotide sequence encoding TANGO 286 with AL021937, however, indicated the presence in TANGO 286 of exons which differ from those disclosed in L021937 (pam120.mat scoring matrix; gap penalties −12/−4). This region of chromosome 22 comprises an immunoglobulin lambda chain C (IGLC) pseudogene, the Ret finger protein-like 3 (RFPL3) and Ret finger protein-like 3 antisense (RFPL3S) genes, a gene encoding a novel immunoglobulin lambda chain V family protein, a novel gene encoding a protein similar both to mouse RGDS protein (RALGDS, RALGEF, guanine nucleotide dissociation stimulator A) and to rabbit oncogene RSC, a novel gene encoding the human orthologue of worm F16A11.2 protein, a novel gene encoding a protein similar both to BPI and to rabbit liposaccharide-binding protein, and a 5′-portion of a novel gene. This region also comprises various ESTs, STSs, GSSs, genomic marker D22S1175, a ca repeat polymorphism and putative CpG islands. TANGO 286 protein thus shares one or more structural or functional features of these molecules.

TANGO 286 protein exhibits considerable sequence similarity with BPI protein, having 23.9% amino acid sequence identity therewith, as assessed using the ALIGN v. 2.0 computer software using a pam120.mat scoring matrix and gap penalties of −12/−4. TANGO 286 protein also exhibits considerable sequence similarity with recombinant endotoxin neutralizing polypeptide (RENP), having 24.5% amino acid sequence identity therewith, as assessed using the ALIGN software. Physiological activities of BPI protein and RENP have been described (e.g., Gabay et al., (1989) Proc. Natl. Acad. Sci. USA 86:5610-5614; Elsbach, (1998) J. Leukoc. Biol. 64:14-18; Mahadeva et al., (1997) Chest 112:1699-1701; International patent application WO96/34873). RENP, for example, binds LPS and neutralizes bacterial endotoxins. BPI, RENP, and other proteins in which a lipid-binding serum glycoprotein domain occurs bind LPS and neutralize bacterial endotoxins, and are therefore useful for preventing, detecting, and treating LPS-related disorders such as shock, disseminated intravascular coagulation, anemia, thrombocytopenia, adult respiratory distress syndrome, renal failure, liver disease, and disorders associated with Gram negative bacterial infections. In addition to the physiological conditions described above, BPI protein is known to be involved in vasculitis and bronchiectasis, in that antibodies which bind specifically with BPI protein are present in at least some patients afflicted with these disorders (Mahadeva et al., supra).

Biological Function of TANGO 286 Proteins, Nucleic Acids, and Modulators Thereof

Expression of TANGO 286 in keratinocyte library indicates that this protein is involved in a disorders which involve keratinocytes. Such disorders include, for example, disorders involving extracellular matrix abnormalities, dermatological disorders, ocular disorders, inappropriate hair growth (e.g., baldness), infections of the nails of the fingers and toes, scalp disorders (e.g., dandruff), and the like.

The fact that TANGO 286 protein contains a lipid-binding serum glycoprotein domain indicates that TANGO 286 is involved in one or more physiological processes in which these other lipid-binding serum glycoprotein domain-containing proteins are involved. Thus, TANGO 286 is involved in one or more of lipid transport, metabolism, serum lipid particle regulation, host anti-microbial defensive mechanisms, and the like.

Human TANGO 286 shares physiological functionality with other proteins in which a lipid-binding serum glycoprotein domains occurs (e.g., LBP, BPI protein, CETP, and PLTP). Based on the amino acid sequence similarity of TANGO 286 with BPI protein and with RENP, TANGO 286 protein exhibits physiological activities exhibited by these proteins. Thus, TANGO 286 proteins are useful for preventing, diagnosing, and treating, among others, lipid transport disorders, lipid metabolism disorders, disorders of serum lipid particle regulation, obesity, disorders involving insufficient or inappropriate host anti-microbial defensive mechanisms, vasculitis, bronchiectasis, LPS-related disorders such as shock, disseminated intravascular coagulation, anemia, thrombocytopenia, adult respiratory distress syndrome, renal failure, liver disease, and disorders associated with Gram negative bacterial infections, such as bacteremia, endotoxemia, sepsis, and the like.

TANGO 294

A cDNA clone (designated jthrc145g07) encoding at least a portion of human TANGO 294 protein was isolated from a human pulmonary artery smooth muscle cell cDNA library. The human TANGO 294 protein is predicted by structural analysis to be a transmembrane membrane protein. No expression of DNA encoding TANGO 294 was detected in human heart, brain, placenta, lung, liver, skeletal muscle, kidney, or pancreas tissues.

The full length of the cDNA encoding TANGO 294 protein (FIG. 6; SEQ ID NO: 45) is 2044 nucleotide residues. The ORF of this cDNA, nucleotide residues 126 to 1394 of SEQ ID NO: 45 (i.e., SEQ ID NO: 46), encodes a 423-amino acid transmembrane protein (FIG. 6; SEQ ID NO: 47).

The invention includes purified TANGO 294 protein, both in the form of the immature 423 amino acid residue protein (SEQ ID NO: 47) and in the form of the mature 390 amino acid residue protein (SEQ ID NO: 49). Mature TANGO 294 protein can be synthesized without the signal sequence polypeptide at the amino terminus thereof, or it can be synthesized by generating immature TANGO 294 protein and cleaving the signal sequence therefrom.

In addition to full length mature and immature TANGO 294 proteins, the invention includes fragments, derivatives, and variants of TANGO 294 protein, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence listed in SEQ ID NO: 45 or some portion thereof, such as the portion which encodes mature TANGO 294 protein, immature .TANGO 294 protein, or a domain of TANGO 294 protein. These nucleic acids are collectively referred to as nucleic acids of the invention.

TANGO 294 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features.

Also included within the scope of the invention are TANGO 294 proteins having a signal sequence. As used herein, a signal sequence includes a peptide of at least about 10 amino acid residues in length which occurs at the amino terminus of membrane-bound proteins and which contains at least about 45% hydrophobic amino acid residues such as alanine, leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferred embodiment, a signal sequence contains at least about 10 to 35 amino acid residues, preferably about 10 to 20 amino acid residues, and has at least about 35-60%, more preferably 40-50%, and more preferably at least about 45% hydrophobic residues. A signal sequence serves to direct a protein containing such a sequence to a lipid bilayer. Thus, in one embodiment, a TANGO 294 protein contains a signal sequence corresponding to amino acid residues 1 to 33 of SEQ ID NO: 47 (SEQ ID NO: 48). The signal sequence is cleaved during processing of the mature protein.

The naturally-occurring form of TANGO 294 protein is a secreted protein (i.e., not comprising the predicted signal sequence). However, in variant forms, TANGO 294 proteins can be transmembrane proteins which include an extracellular domain. In this transmembrane variant form, the predicted TANGO 294 protein extracellular domain is located from about amino acid residue 34 to about amino acid residue 254 of SEQ ID NO: 47, the predicted cytoplasmic domain is located from about amino acid residue 280 to amino acid residue 423 of SEQ ID NO: 47 (SEQ ID NO: 52), and the predicted transmembrane domain is located from about amino acid residues 255 to 279 of SEQ ID NO: 47 (SEQ ID NO: 51).

TANGO 294 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Table X, as predicted by computerized sequence analysis of TANGO 294 proteins using amino acid sequence comparison software (comparing the amino acid sequence of TANGO 294 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2, 4, 6, 10, 15, or 20 or more of the post-translational modification sites listed in Table X.

TABLE X Type of Potential Modification Amino Acid Residues Amino Acid Site or Domain of SEQ ID NO: 47 Sequence N-glycosylation site 48 to 51 NISE 113 to 116 NNSL 285 to 288 NMSR 413 to 416 NLSQ Protein kinase C 12 to 14 SHR phosphorylation site 138 to 140 SRK 217 to 219 TVK Casein kinase II 155 to 158 SYDE phosphorylation site 175 to 178 TGQE 198 to 201 TMPE 360 to 363 SNPE Tyrosine kinase 174 to 182 KTGQEKIYY phosphorylation site N-myristoylation site 99 to 104 GLVGGA 130 to 135 GNSRGN 188 to 193 GTTMGF 277 to 282 GGFNTN Amidation site 240 to 243 FGKK Lipase serine active site 180 to 189 IYYVGYSQGT Alpha/beta hydrolase 125 to 404 See FIG. 6 fold domain

Alpha/beta hydrolase fold domains occur in a wide variety of enzymes (Ollis et al., (1992) Protein Eng. 5:197-211). The alpha/beta fold domain is a conserved topological domain in which sequence homology is not necessarily conserved. Conservation of topology in the alpha/beta fold domain preserves arrangement of catalytic residues, even though those residues, and the reactions they catalyze, can vary. In many enzymes, particularly including alpha/beta hydrolases, this domain encompasses the active site of the enzyme. In one embodiment, the protein of the invention has at least one domain that is at least 55%, preferably at least about 65%, more preferably at least about 75%, yet more preferably at least about 85%, and most preferably at least about 95% identical to the alpha/beta hydrolase fold domain described herein in Table X.

The signal peptide prediction program SIGNALP (Nielsen et al. (1997) Protein Engineering 10:1-6) predicted that human TANGO 294 protein includes a 33 amino acid signal peptide (amino acid residues 1 to 33 of SEQ ID NO: 47; SEQ ID NO: 48) preceding the mature TANGO 294 protein (amino acid residues 34 to 423 of SEQ ID NO: 47; SEQ ID NO: 49). Human TANGO 294 protein is a soluble secreted protein. However, in the transmembrane variant form, human TANGO 294 protein includes an extracellular domain (amino acid residues 34 to 254 of SEQ ID NO: 47; SEQ ID NO: 50); a transmembrane domain (amino acid residues 255 to 279 of SEQ ID NO: 47; SEQ ID NO: 51), and a cytoplasmic domain (amino acid residues 280 to 423 of SEQ ID NO: 47; SEQ ID NO: 52)

FIG. 6F depicts a hydrophilicity plot of human TANGO 294 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic region which corresponds to amino acid residues 1 to 33 of SEQ ID NO: 47 is the signal sequence of human TANGO 294 (SEQ ID NO: 49). The hydrophobic region which corresponds to amino acid residues 255 to 279 of SEQ ID NO: 47 is the predicted transmembrane domain of human TANGO 294 (SEQ ID NO: 51). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human TANGO 294 protein from about amino acid residue 130 to about amino acid residue 150 appears to be located at or near the surface of the protein, while the region from about amino acid residue 90 to about amino acid residue 100 appears not to be located at or near the surface.

The predicted molecular weight of human TANGO 294 protein without modification and prior to cleavage of the signal sequence is about 48.2 kilodaltons. The predicted molecular weight of the mature human TANGO 294 protein without modification and after cleavage of the signal sequence is about 44.2 kilodaltons.

It may be that amino acid residues 1 to 15 of SEQ ID NO: 47 do not occur in TANGO 294 protein. However, it is recognized that amino acid residues 16 to 33 of SEQ ID NO: 47 form a functional signal sequence even in the absence of residues 1 to 15. The amino acid sequence (and hence the properties) of mature TANGO 294 protein are unaffected by presence or absence of amino acid residues 1 to 15 of immature TANGO 294 protein.

Human TANGO 294 protein exhibits considerable sequence similarity (i.e., about 75% amino acid sequence identity) to lingual and gastric lipase proteins of rat (Swissprot Accession no. P04634; Docherty et al. (1985) Nucleic Acids Res. 13:1891-1903), dog (Swissprot Accession no. P80035; Carriere et al. (1991) Eur. J. Biochem. 202:75-83), and human (Swissprot Accession no. P07098; Bernbaeck and Blaeckberg (1987) Biochim. Biophys. Acta 909:237-244), as assessed using the ALIGN v.2.0 computer software using a pam12.mat scoring matrix and gap penalties of −12/−4. TANGO 294 is distinct from the known human lipase, as indicated in FIGS. 6D and 6E. FIGS. 6D and 6E depict an alignment of the amino acid sequences of human TANGO 294 protein (SEQ ID NO: 47) and the known human lipase protein (SEQ ID NO: 75), as assessed using the same software and parameters. In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the amino acid sequences of the proteins are 49.8% identical. TANGO 294 also is distinct from the known human lysosomal acid lipase, as indicated in FIGS. 6G and 6H. FIGS. 6G and 6H depicts an alignment of the amino acid sequences of human TANGO 294 protein (SEQ ID NO: 47) and the known human lysosomal acid lipase protein (SEQ ID NO: 41). In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the amino acid sequences of the proteins are 56.9% identical.

TANGO 294 is a human lipase distinct from the known human lipase and the known human lysosomal acid lipase. Furthermore, in view of the comparisons of the amino acid sequences of TANGO 294 and the two human lipases and the nature of transcriptional initiation sites, it is recognized that the transcriptional start site can correspond to either of the methionine residues located at residues 1 and 15 of SEQ ID NO: 47 The present invention thus includes proteins in which the initially transcribed amino acid residue is the methionine residue at position 1 of SEQ ID NO: 47 and proteins in which the initially transcribed amino acid residue is the methionine residue at position 15 of SEQ ID NO: 47 (i.e., proteins in which the amino acid sequence of TANGO 294 does not include residues 1 to 14 of SEQ ID NO: 47). Furthermore, because amino acid residues 1 to 14 of SEQ ID NO: 47 are predicted to be part of a signal sequence, it is recognized that the protein not comprising this portion of the amino acid sequence will nonetheless exhibit a functional signal sequence at its amino terminus.

Biological Function of TANGO 294 Proteins, Nucleic Acids, and Modulators Thereof

The sequence similarity of TANGO 294 and mammalian lingual, gastric, and lysosomal acid lipase proteins indicates that TANGO 294 is involved in physiological processes identical or analogous to those involving these lipases. Thus, TANGO 294 is involved in facilitating absorption and metabolism of fat. TANGO 294 can thus be used, for example, to prevent, detect, and treat disorders relating to fat absorption and metabolism, such as inadequate expression of gastric/pancreatic lipase, cystic fibrosis, exocrine pancreatic insufficiency, obesity, medical treatments which alter fat absorption, and the like.

TANGO 294 protein is known to be expressed in human pulmonary artery smooth muscle tissue. This indicates that TANGO 294 protein is involved in transportation and metabolism of fats and lipids in the human vascular and cardiovascular systems. Thus, TANGO 294 proteins of the invention can be used to prevent, detect, and treat disorders involving these body systems.

INTERCEPT 296

A cDNA clone (designated jthEa030h09) encoding at least a portion of human INTERCEPT 296 protein was isolated from a human esophagus cDNA library. The human INTERCEPT 296 protein is predicted by structural analysis to be a transmembrane protein having three or more transmembrane domains. Expression of DNA encoding INTERCEPT 296 tissue has been detected by northern analysis of human lung tissue. In human lung tissue, two moieties corresponding to INTERCEPT 296 have been identified in Northern blots. It is recognized that these two moieties may represent alternatively polyadenylated INTERCEPT 296 mRNAs or alternatively spliced INTERCEPT 296 mRNAs. It has furthermore been observed that INTERCEPT 296 does not appear to be expressed in any of heart, brain, placenta, skeletal muscle, kidney, and pancreas tissues.

The full length of the cDNA encoding INTERCEPT 296 protein (FIG. 7; SEQ ID NO: 53) is 2133 nucleotide residues. The ORF of this cDNA, nucleotide residues 70 to 1098 of SEQ ID NO: 53 (i.e., SEQ ID NO: 54), encodes a 343-amino acid transmembrane protein (FIG. 7; SEQ ID NO: 55).

The invention includes purified INTERCEPT 296 protein, which has the amino acid sequence listed in SEQ ID NO: 55. In addition to full length INTERCEPT 296 proteins, the invention includes fragments, derivatives, and variants of these INTERCEPT 296 proteins, as described herein. These proteins, fragments, derivatives, and variants are collectively referred to herein as polypeptides of the invention or proteins of the invention.

The invention also includes nucleic acid molecules which encode a polypeptide of the invention. Such nucleic acids include, for example, a DNA molecule having the nucleotide sequence SEQ ID NO: 53 or some portion thereof, such as the portion which encodes INTERCEPT 296 protein or a domain thereof. These nucleic acids are collectively referred to as nucleic acids of the invention.

INTERCEPT 296 proteins and nucleic acid molecules encoding them comprise a family of molecules having certain conserved structural and functional features, such as the five transmembrane domains which occur in the protein.

INTERCEPT 296 comprises at least five transmembrane domains, at least three cytoplasmic domains, and at least two extracellular domains. INTERCEPT 296 does not appear to comprise a cleavable signal sequence. Amino acid residues 1 to 70 of SEQ ID NO: 55 likely directs insertion of the protein into the cytoplasmic membrane. There are at least two mechanisms by which this can occur. Sequence analysis of residues 1 to 70 of SEQ ID NO: 55 indicates that this entire region may represent a signal sequence or that residues 1 to 47 represent a signal sequence, with residues 48-70 representing a transmembrane region. Human INTERCEPT 296 protein extracellular domains are located from about amino acid residue 70 to about amino acid residue 182 (SEQ ID NO: 57) and from about amino acid residue 228 to about amino acid residue 249 (SEQ ID NO: 58) of SEQ ID NO: 55. Human INTERCEPT 296 cytoplasmic domains are located from about amino acid residue 43 to amino acid residue 50 (SEQ ID NO: 64), from about amino acid residue 205 to amino acid residue 210 (SEQ ID NO: 65), and from amino acid residue 272 to amino acid residue 343 (SEQ ID NO: 66) of SEQ ID NO: 55. The five transmembrane domains of INTERCEPT 296 are located from about amino acid residues 24 to 42 (SEQ ID NO: 59), 51 to 70 (SEQ ID NO: 60), 183 to 204 (SEQ ID NO: 61), 211 to 227 (SEQ ID NO: 62), and 250 to 271 (SEQ ID NO: 63) of SEQ ID NO: 55.

INTERCEPT 296 proteins typically comprise a variety of potential post-translational modification sites (often within an extracellular domain), such as those described herein in Table XI, as predicted by computerized sequence analysis of INTERCEPT 296 proteins using amino acid sequence comparison software (comparing the amino acid sequence of INTERCEPT 296 with the information in the PROSITE database {rel. 12.2; February 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certain embodiments, a protein of the invention has at least 1, 2,4, 6, 10, 15, or 20 or more of the post-translational modification sites listed in Table XI.

TABLE XI Type of Potential Modification Amino Acid Residues Amino Acid Site or Domain of SEQ ID NO: 55 Sequence N-glycosylation site 71 to 74 NFSS 84 to 87 NTSY 109 to 112 NITL 121 to 124 NETI 284 to 287 NQSV Protein kinase C 86 to 88 SYK phosphorylation site 131 to 133 TWR 162 to 164 TPR 304 to 306 SPR 313 to 315 SPK 326 to 328 STK Casein kinase II 286 to 289 SVDE phosphorylation site 296 to 299 SPEE 309 to 312 SMAD Tyrosine kinase 148 to 156 KGLPDPVLY phosphorylation site N-myristoylation site 79 to 84 GQVSTN 100 to 105 GLQVGL 107 to 112 GVNITL 265 to 270 GLAMAV

FIG. 7D depicts a hydrophilicity plot of INTERCEPT 296 protein. Relatively hydrophobic regions are above the dashed horizontal line, and relatively hydrophilic regions are below the dashed horizontal line. The hydrophobic regions which corresponds to amino acid residues 24 to 42, 51 to 70, 183 to 204, 211 to 227, and 250 to 271 of SEQ ID NO: 55 are the transmembrane domains of human INTERCEPT 296 (SEQ ID NOs: 59 through 63, respectively). As described elsewhere herein, relatively hydrophilic regions are generally located at or near the surface of a protein, and are more frequently effective immunogenic epitopes than are relatively hydrophobic regions. For example, the region of human INTERCEPT 296 protein from about amino acid residue 120 to about amino acid residue 140 appears to be located at or near the surface of the protein, while the region from about amino acid residue 95 to about amino acid residue 110 appears not to be located at or near the surface.

The predicted molecular weight of INTERCEPT 296 protein without modification and prior to cleavage of the signal sequence is about 37.8 kilodaltons. The predicted molecular weight of the mature INTERCEPT 296 protein without modification and after cleavage of the signal sequence is about 30.2 kilodaltons.

FIGS. 7E and 7F depicts an alignment of the amino acid sequences of human INTERCEPT 296 protein (SEQ ID NO: 55) and Caenorhabditis elegans C06E1.3 related protein (SEQ ID NO: 399). In this alignment (pam120.mat scoring matrix, gap penalties −12/−4), the amino acid sequences of the proteins are 26.8% identical. The C. elegans protein has five predicted transmembrane domains.

Biological Function of INTERCEPT 296 Proteins, Nucleic Acids, and Modulators Thereof

The cDNA encoding INTERCEPT 296 protein was obtained from a human esophagus cDNA library, and INTERCEPT 296 is expressed in lung tissue. The INTERCEPT 296-related proteins and nucleic acids of the invention are therefore useful for prevention, detection, and treatment of disorders of the human lung and esophagus. Such disorders include, for example, various cancers, bronchitis, cystic fibrosis, respiratory infections (e.g., influenza, bronchiolitis, pneumonia, and tuberculosis), asthma, emphysema, chronic bronchitis, bronchiectasis, pulmonary edema, pleural effusion, pulmonary embolus, adult and infant respiratory distress syndromes, heartburn, and gastric reflux esophageal disease.

Tables A and B summarize sequence data corresponding to the human proteins herein designated TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296.

TABLE A Protein SEQ ID NOs Depicted in ATCC ® Designation cDNA ORF Protein FIG. # Accession # TANGO 202  1  2  3 1 207219 TANGO 234  9 10 11 2 207184 TANGO 265 17 18 19 3 207228 TANGO 273 25 26 27 4 207185 TANGO 286 33 34 35 5 207220 TANGO 294 45 46 47 6 207220 INTERCEPT 296 53 54 55 7 207220

TABLE B Extracellular Transmembrane Cytoplasmic Protein Desig. Signal Sequence Mature Protein Domain(s) Domain(s) Domain(s) SEQ ID NOs TANGO 202 1 to 19  4 20 to 475  5 20 to 392  6 393 to 415  7 416 to 475  8 (variant) (1 to 19)  (4) (20 to 475)  (5) (20 to 475)  (5) (N/A) (N/A) TANGO 234 1 to 40 12  41 to 1453 13  41 to 1359 14 1360 to 1383 15 1384 to 1453 16 TANGO 265 1 to 31 20 32 to 761 21 32 to 683 22 684 to 704 23 705 to 761 24 TANGO 273 1 to 22 28 23 to 172 29 23 to 60  30 61 to 81 31  82 to 172 32 TANGO 286 1 to 23 36 24 to 455 37 24 to 455 37 N/A N/A TANGO 294 1 to 33 48 34 to 423 49 34 to 254 50 255 to 279 51 280 to 423 52 (variant 1) (15 to 33)  (40) (34 to 423) (49) (34 to 254) (50) (255 to 279) (51) (280 to 423) (52) <variant 2> <1 to 33> <48> <34 to 423> <49> <34 to 423> <49> <N/A> <N/A> {variant 3} {15 to 33}  {40} {34 to 423} {49} {34 to 423} {49} {N/A} {N/A} INTERCEPT N/A  1 to 343 55 1 to 23 56 24 to 42 59 43 to 50 64 296 71 to 182 57 51 to 70 60 205 to 210 65 228 to 249  58 183 to 204 61 272 to 343 66 211 to 227 62 250 to 271 63 Amino Acid Residues

Various aspects of the invention are described in further detail in the following subsections.

I. Isolated Nucleic Acid Molecules

One aspect of the invention pertains to isolated nucleic acid molecules that encode a polypeptide of the invention or a biologically active portion thereof, as well as nucleic acid molecules sufficient for use as hybridization probes to identify nucleic acid molecules encoding a polypeptide of the invention and fragments of such nucleic acid molecules suitable for use as PCR primers for the amplification or mutation of nucleic acid molecules. As used herein, the term “nucleic acid molecule” is intended to include DNA molecules (e.g., cDNA or genomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.

An “isolated” nucleic acid molecule is one which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid molecule. Preferably, an “isolated” nucleic acid molecule is free of sequences (preferably protein-encoding sequences) which naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated nucleic acid molecule can contain less than about 5 kB, 4 kB, 3 kB, 2 kB, 1 kB, 0.5 kB or 0.1 kB of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized.

A nucleic acid molecule of the present invention, e.g., a nucleic acid molecule having the nucleotide sequence of all or a portion of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a complement thereof, or which has a nucleotide sequence comprising one of these sequences, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using a nucleic acid comprising at least one of the sequences of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 as a hybridization probe, nucleic acid molecules of the invention can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook et al., eds., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).

A nucleic acid molecule of the invention can be amplified using cDNA, mRNA or genomic DNA as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to all or a portion of a nucleic acid molecule of the invention can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.

In another preferred embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule which is a complement of the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a portion thereof. A nucleic acid molecule which is complementary to a given nucleotide sequence is one which is sufficiently complementary to the given nucleotide sequence that it can hybridize to the given nucleotide sequence thereby forming a stable duplex.

Moreover, a nucleic acid molecule of the invention can comprise only a portion of a nucleic acid sequence encoding a full length polypeptide of the invention for example, a fragment which can be used as a probe or primer or a fragment encoding a biologically active portion of a polypeptide of the invention. The nucleotide sequence determined from the cloning one gene allows for the generation of probes and primers designed for use in identifying and/or cloning homologs in other cell types, e.g., from other tissues, as well as homologs from other mammals. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 15, preferably about 25, more preferably about 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, or 400 or more consecutive nucleotides of the sense or anti-sense sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or of a naturally occurring mutant of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73.

Probes based on the sequence of a nucleic acid molecule of the invention can be used to detect transcripts or genomic sequences encoding the same protein molecule encoded by a selected nucleic acid molecule. The probe comprises a label group attached thereto, e.g., a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as part of a diagnostic test kit for identifying cells or tissues which mis-express the protein, such as by measuring levels of a nucleic acid molecule encoding the protein in a sample of cells from a subject, e.g., detecting mRNA levels or determining whether a gene encoding the protein has been mutated or deleted.

A nucleic acid fragment encoding a biologically active portion of a polypeptide of the invention can be prepared by isolating a portion of any of SEQ ID NOs: 2, 10, 18, 26, 34, 46, 54, 68, and 73, expressing the encoded portion of the polypeptide protein (e.g., by recombinant expression in vitro), and assessing the activity of the encoded portion of the polypeptide.

The invention further encompasses nucleic acid molecules that differ from the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 due to degeneracy of the genetic code and thus encode the same protein as that encoded by the nucleotide sequence of any of SEQ ID NOs: 2, 10, 18, 26, 34, 46, 54, 68, and 73.

In addition to the nucleotide sequences of SEQ ID NOs: 2, 10, 18, 26, 34, 46, 54, 68, and 73, it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequence can exist within a population (e.g., the human population). Such genetic polymorphisms can exist among individuals within a population due to natural allelic variation. An allele is one of a group of genes which occur alternatively at a given genetic locus.

As used herein, the phrase “allelic variant” refers to a nucleotide sequence which occurs at a given locus or to a polypeptide encoded by the nucleotide sequence. For example, chromosomal mapping has been used to locate the gene encoding human TANGO 234 at chromosomal location h12p13 (with synteny to mo6), between chromosomal markers WI-6980 and GATA8A09.43. Thus, human TANGO 234 allelic variants can include TANGO 234 nucleotide sequence polymorphisms (e.g., nucleotide sequences that vary from SEQ ID NO: 9) that map to this chromosomal region. Similarly, chromosomal mapping has been used to locate the gene encoding human TANGO 265 protein on chromosome 1, between markers D1S305 and D1S2635. Allelic variants of TANGO 265 occur at this chromosomal location. Further by way of example, the gene encoding human TANGO 273 protein has been located by chromosomal mapping on chromosome 7, between markers D7S2467 and D7S2552. Allelic variants of TANGO 273 occur at this chromosomal location.

As used herein, the terms “gene” and “recombinant gene” refer to nucleic acid molecules comprising an open reading frame encoding a polypeptide of the invention. Such natural allelic variations can typically result in 1-5% variance in the nucleotide sequence of a given gene. Alternative alleles can be identified by sequencing the gene of interest in a number of different individuals. This can be readily carried out by using hybridization probes to identify the same genetic locus in a variety of individuals. Any and all such nucleotide variations and resulting amino acid polymorphisms or variations that are the result of natural allelic variation and that do not alter the functional activity are intended to be within the scope of the invention.

Moreover, nucleic acid molecules encoding proteins of the invention from other species (homologs), which have a nucleotide sequence which differs from that of the specific proteins described herein are intended to be within the scope of the invention. Nucleic acid molecules corresponding to natural allelic variants and homologs of a cDNA of the invention can be isolated based on their homology with nucleic acid molecules described herein, using the specific cDNAs described herein, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions. For example, a cDNA encoding a soluble form of a membrane-bound protein of the invention isolated based on its hybridization to a nucleic acid molecule encoding all or part of the membrane-bound form. Likewise, a cDNA encoding a membrane-bound form can be isolated based on its hybridization to a nucleic acid molecule encoding all or part of the soluble form.

Accordingly, in another embodiment, an isolated nucleic acid molecule of the invention is at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300, 350, 400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3500, 4000, 4500, or 4928) nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence, preferably the coding sequence, of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a complement thereof. As used herein, the term “hybridizes-under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% (65%, 70%, preferably 75%) identical to each other typically remain hybridized to each other. Such stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. A preferred, non-limiting example of stringent hybridization conditions are hybridization in 6×sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65° C. Preferably, an isolated nucleic acid molecule of the invention that hybridizes under stringent conditions with the sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a complement thereof, corresponds to a naturally-occurring nucleic acid molecule. As used herein, a “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).

In addition to naturally-occurring allelic variants of a nucleic acid molecule of the invention sequence that can exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation thereby leading to changes in the amino acid sequence of the encoded protein, without altering the biological activity of the protein. For example, one can make nucleotide substitutions leading to amino acid substitutions at “non-essential” amino acid residues. A “non-essential” amino acid residue is a residue that can be altered from the wild-type sequence without altering the biological activity, whereas an “essential” amino acid residue is required for biological activity. For example, amino acid residues that are not conserved or only semi-conserved among homologs of various species may be non-essential for activity and thus would be likely targets for alteration. Alternatively, amino acid residues that are conserved among the homologs of various species (e.g., murine and human) may be essential for activity and thus would not be likely targets for alteration.

Accordingly, another aspect of the invention pertains to nucleic acid molecules encoding a polypeptide of the invention that contain changes in amino acid residues that are not essential for activity. Such polypeptides differ in amino acid sequence from the sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, yet retain biological activity. In one embodiment, the isolated nucleic acid molecule includes a nucleotide sequence encoding a protein that includes an amino acid sequence that is at least about 40% identical, 50%, 60%, 70%, 80%, 90%, 95%, or 98% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74.

An isolated nucleic acid molecule encoding a variant protein can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, such that one or more amino acid residue substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Alternatively, mutations can be introduced randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed recombinantly and the activity of the protein can be determined.

In a preferred embodiment, a mutant polypeptide that is a variant of a polypeptide of the invention can be assayed for: (1) the ability to form protein:protein interactions with one or more polypeptides of the invention (e.g., in a signaling pathway); (2) the ability to bind a ligand of a polypeptide of the invention (e.g., another protein identified herein); (3) the ability to bind to an intracellular target protein of a polypeptide of the invention (e.g., a modulator or substrate of the polypeptide); or (4) the ability to modulate a physiological activity of the protein, such as one of those disclosed herein (e.g., ability to modulate cell proliferation, cell migration, chemotaxis, or cellular differentiation).

The present invention encompasses antisense nucleic acid molecules, i.e., molecules which are complementary to a sense nucleic acid encoding a polypeptide of the invention, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. Accordingly, an antisense nucleic acid can hydrogen bond to a sense nucleic acid. The antisense nucleic acid can be complementary to an entire coding strand, or to only a portion thereof, e.g., all or part of the protein coding region (or open reading frame). An antisense nucleic acid molecule can be antisense to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a polypeptide of the invention. The non-coding regions (“5′ and 3′ untranslated regions”) are the 5′ and 3′ sequences which flank the coding region and are not translated into amino acids.

An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 or more nucleotides in length. An antisense nucleic acid of the invention can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. Examples of modified nucleotides which can be used to generate the antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N₆-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been sub-cloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).

The antisense nucleic acid molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a selected polypeptide of the invention to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule which binds to DNA duplexes, through specific interactions in the major groove of the double helix. An example of a route of administration of antisense nucleic acid molecules of the invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface, e.g., by linking the antisense nucleic acid molecules to peptides or antibodies which bind to cell surface receptors or antigens. The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient intracellular concentrations of the antisense molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.

An antisense nucleic acid molecule of the invention can be an alpha-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual beta-units, the strands run parallel to each other (Gaultier et al. (1987) Nucleic Acids Res. 15:6625-6641). The antisense nucleic acid molecule can also comprise a 2′-o-methylribonucleotide (Inoue et al. (1987) Nucleic Acids Res. 15:6131-6148) or a chimeric RNA-DNA analogue (Inoue et al. (1987) FEBS Lett. 215:327-330).

The invention also encompasses ribozymes. Ribozymes are catalytic RNA molecules with ribonuclease activity which are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes as described in Haselhoff and Gerlach (1988) Nature 334:585-591) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a nucleic acid molecule encoding a polypeptide of the invention can be designed based upon the nucleotide sequence of a cDNA disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a Cech et al. U.S. Pat. No. 4,987,071; and Cech et al. U.S. Pat. No. 5,116,742. Alternatively, an mRNA encoding a polypeptide of the invention can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel and Szostak (1993) Science 261:1411-1418.

The invention also encompasses nucleic acid molecules which form triple helical structures. For example, expression of a polypeptide of the invention can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the polypeptide (e.g., the promoter and/or enhancer) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene (1991) Anticancer Drug Des. 6(6):569-84; Helene (1992) Ann N.Y. Acad. Sci. 660:27-36; and Maher (1992) Bioassays 14(12):807-15.

In various embodiments, the nucleic acid molecules of the invention can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see Hyrup et al. (1996) Bioorganic & Medicinal Chemistry 4(1): 5-23). As used herein, the terms “peptide nucleic acids” or “PNAs” refer to nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup et al. (1996), supra; Perry-O'Keefe et al. (1996) Proc. Natl. Acad. Sci. USA 93: 14670-675.

PNAs can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or anti-gene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs can also be used, e.g., in the analysis of single base pair mutations in a gene by, e.g., PNA directed PCR clamping; as artificial restriction enzymes when used in combination with other enzymes, e.g., S1 nucleases (Hyrup (1996), supra; or as probes or primers for DNA sequence and hybridization (Hyrup (1996), supra; Perry-O'Keefe et al. (1996) Proc. Natl. Acad. Sci. USA 93: 14670-675).

In another embodiment, PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which can combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNase H and DNA polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (Hyrup (1996), supra). The synthesis of PNA-DNA chimeras can be performed as described in Hymp (1996), supra, and Finn et al. (1996) Nucleic Acids Res. 24(17):3357-63. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5′-(4-methoxytrityl)amino-5′-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5′ end of DNA (Mag et al. (1989) Nucleic Acids Res. 17:5973-88). PNA monomers are then coupled in a step-wise manner to produce a chimeric molecule with a 5′ PNA segment and a 3′ DNA segment (Finn et al. (1996) Nucleic Acids Res. 24(17):3357-63). Alternatively, chimeric molecules can be synthesized with a 5′ DNA segment and a 3′ PNA segment (Peterser et al. (1975) Bioorganic Med. Chem. Lett. 5:1119-11124).

In other embodiments, the oligonucleotide can include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger et al. (1989) Proc. Natl. Acad. Sci. USA 86:6553-6556; Lemaitre et al. (1987) Proc. Natl. Acad. Sci. USA 84:648-652; PCT Publication No. WO 88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134). In addition, oligonucleotides can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al. (1988) Bio/Techniques 6:958-976) or intercalating agents (see, e.g., Zon (1988) Pharm. Res. 5:539-549). To this end, the oligonucleotide can be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.

II. Isolated Proteins and Antibodies

One aspect of the invention pertains to isolated proteins, and biologically active portions thereof, as well as polypeptide fragments suitable for use as immunogens to raise antibodies directed against a polypeptide of the invention. In one embodiment, the native polypeptide can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. In another embodiment, polypeptides of the invention are produced by recombinant DNA techniques. Alternative to recombinant expression, a polypeptide of the invention can be synthesized chemically using standard peptide synthesis techniques.

An “isolated” or “purified” protein or biologically active portion thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or substantially free of chemical precursors or other chemicals when chemically synthesized. The language “substantially free of cellular material” includes preparations of protein in which the protein is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, or 5% (by dry weight) of heterologous protein (also referred to herein as a “contaminating protein”). When the protein or biologically active portion thereof is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, or 5% of the volume of the protein preparation. When the protein is produced by chemical synthesis, it is preferably substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals which are involved in the synthesis of the protein. Accordingly such preparations of the protein have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than the polypeptide of interest.

Biologically active portions of a polypeptide of the invention include polypeptides comprising amino acid sequences sufficiently identical to or derived from the amino acid sequence of the protein (e.g., the amino acid sequence shown in any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74), which include fewer amino acids than the full length protein, and exhibit at least one activity of the corresponding full-length protein. Typically, biologically active portions comprise a domain or motif with at least one activity of the corresponding protein. A biologically active portion of a protein of the invention can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acids in length. Moreover, other biologically active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the functional activities of the native form of a polypeptide of the invention.

Preferred polypeptides have the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74. Other useful proteins are substantially identical (e.g., at least about 40%, preferably 50%, 60%, 70%, 80%, 90%, 95%, or 99%) to any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74 and retain the functional activity of the protein of the corresponding naturally-occurring protein yet differ in amino acid sequence due to natural allelic variation or mutagenesis.

To determine the percent identity of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=# of identical positions/total # of positions (e.g., overlapping positions)×100). In one embodiment the two sequences are the same length.

The determination of percent identity between two sequences can be accomplished using a mathematical algorithm. A preferred, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul, et al. (1990) J. Mol. Biol. 215:403-410. BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecules of the invention. BLAST protein searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to a protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402. Alternatively, PSI-Blast can be used to perform an iterated search which detects distant relationships between molecules. Id. When utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http://www.ncbi.nlm.nih.gov. Another preferred, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, (1988) CABIOS 4:11-17. Such an algorithm is incorporated into the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, only exact matches are counted.

The invention also provides chimeric or fusion proteins. As used herein, a “chimeric protein” or “fusion protein” comprises all or part (preferably biologically active) of a polypeptide of the invention operably linked to a heterologous polypeptide (i.e., a polypeptide other than the same polypeptide of the invention). Within the fusion protein, the term “operably linked” is intended to indicate that the polypeptide of the invention and the heterologous polypeptide are fused in-frame to each other. The heterologous polypeptide can be fused to the amino-terminus or the carboxyl-terminus of the polypeptide of the invention.

One useful fusion protein is a GST fusion protein in which the polypeptide of the invention is fused to the carboxyl terminus of GST sequences. Such fusion proteins can facilitate the purification of a recombinant polypeptide of the invention.

In another embodiment, the fusion protein contains a heterologous signal sequence at its amino terminus. For example, the native signal sequence of a polypeptide of the invention can be removed and replaced with a signal sequence from another protein. For example, the gp67 secretory sequence of the baculovirus envelope protein can be used as a heterologous signal sequence (Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley & Sons, 1992). Other examples of eukaryotic heterologous signal sequences include the secretory sequences of melittin and human placental alkaline phosphatase (Stratagene; La Jolla, Calif.). In yet another example, useful prokaryotic heterologous signal sequences include the phoA secretory signal (Sambrook et al., supra) and the protein A secretory signal (Pharmacia Biotech; Piscataway, N.J.).

In yet another embodiment, the fusion protein is an immunoglobulin fusion protein in which all or part of a polypeptide of the invention is fused to sequences derived from a member of the immunoglobulin protein family. The immunoglobulin fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between a ligand (soluble or membrane-bound) and a protein on the surface of a cell (receptor), to thereby suppress signal transduction in vivo. The immunoglobulin fusion protein can be used to affect the bioavailability of a cognate ligand of a polypeptide of the invention. Inhibition of ligand/receptor interaction can be useful therapeutically, both for treating proliferative and differentiative disorders and for modulating (e.g., promoting or inhibiting) cell survival. Moreover, the immunoglobulin fusion proteins of the invention can be used as immunogens to produce antibodies directed against a polypeptide of the invention in a subject, to purify ligands and in screening assays to identify molecules which inhibit the interaction of receptors with ligands.

Chimeric and fusion proteins of the invention can be produced by standard recombinant DNA techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and re-amplified to generate a chimeric gene sequence (see, e.g., Ausubel et al., supra). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). A nucleic acid encoding a polypeptide of the invention can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the polypeptide of the invention.

A signal sequence of a polypeptide of the invention (e.g., the signal sequence in one of SEQ ID NOs: 3, 4, 11, 12, 19, 20, 27, 28, 35, 36, 47, 48, 69, and 74) can be used to facilitate secretion and isolation of the secreted protein or other proteins of interest. Signal sequences are typically characterized by a core of hydrophobic amino acids which are generally cleaved from the mature protein during secretion in one or more cleavage events. Such signal peptides contain processing sites that allow cleavage of the signal sequence from the mature proteins as they pass through the secretory pathway. Thus, the invention pertains to the described polypeptides having a signal sequence, as well as to the signal sequence itself and to the polypeptide in the absence of the signal sequence (i.e., the cleavage products). In one embodiment, a nucleic acid sequence encoding a signal sequence of the invention can be operably linked in an expression vector to a protein of interest, such as a protein which is ordinarily not secreted or is otherwise difficult to isolate. The signal sequence directs secretion of the protein, such as from a eukaryotic host into which the expression vector is transformed, and the signal sequence is subsequently or concurrently cleaved. The protein can then be readily purified from the extracellular medium by art recognized methods. Alternatively, the signal sequence can be linked to the protein of interest using a sequence which facilitates purification, such as with a GST domain.

In another embodiment, the signal sequences of the present invention can be used to identify regulatory sequences, e.g., promoters, enhancers, repressors. Since signal sequences are the most amino-terminal sequences of a peptide, it is expected that the nucleic acids which flank the signal sequence on its amino-terminal side will be regulatory sequences which affect transcription. Thus, a nucleotide sequence which encodes all or a portion of a signal sequence can be used as a probe to identify and isolate signal sequences and their flanking regions, and these flanking regions can be studied to identify regulatory elements therein.

The present invention also pertains to variants of the polypeptides of the invention. Such variants have an altered amino acid sequence which can function as either agonists (mimetics) or as antagonists. Variants can be generated by mutagenesis, e.g., discrete point mutation or truncation. An agonist can retain substantially the same, or a subset, of the biological activities of the naturally occurring form of the protein. An antagonist of a protein can inhibit one or more of the activities of the naturally occurring form of the protein by, for example, competitively binding to a downstream or upstream member of a cellular signaling cascade which includes the protein of interest. Thus, specific biological effects can be elicited by treatment with a variant of limited function. Treatment of a subject with a variant having a subset of the biological activities of the naturally occurring form of the protein can have fewer side effects in a subject relative to treatment with the naturally occurring form of the protein.

Variants of a protein of the invention which function as either agonists (mimetics) or as antagonists can be identified by screening combinatorial libraries of mutants, e.g., truncation mutants, of the protein of the invention for agonist or antagonist activity. In one embodiment, a variegated library of variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential protein sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display). There are a variety of methods which can be used to produce libraries of potential variants of the polypeptides of the invention from a degenerate oligonucleotide sequence. Methods for synthesizing degenerate oligonucleotides are known in the art (see, e.g., Narang (1983) Tetrahedron 39:3; Itakura et al. (1984) Annu. Rev. Biochem. 53:323; Itakura et al. (1984) Science 198:1056; Ike et al. (1983) Nucleic Acid Res. 11:477).

In addition, libraries of fragments of the coding sequence of a polypeptide of the invention can be used to generate a variegated population of polypeptides for screening and subsequent selection of variants. For example, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of the coding sequence of interest with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, re-naturing the DNA to form double stranded DNA which can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with S1 nuclease, and ligating the resulting fragment library into an expression vector. By this method, an expression library can be derived which encodes amino terminal and internal fragments of various sizes of the protein of interest.

Several techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. The most widely used techniques, which are amenable to high through-put analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a technique which enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify variants of a protein of the invention (Arkin and Yourvan (1992) Proc. Natl. Acad. Sci. USA 89:7811-7815; Delgrave et al. (1993) Protein Engineering 6(3):327-331).

An isolated polypeptide of the invention, or a fragment thereof, can be used as an immunogen to generate antibodies using standard techniques for polyclonal and monoclonal antibody preparation. The full-length polypeptide or protein can be used or, alternatively, the invention provides antigenic peptide fragments for use as immunogens. The antigenic peptide of a protein of the invention comprises at least 8 (preferably 10, 15, 20, or 30 or more) amino acid residues of the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, and encompasses an epitope of the protein such that an antibody raised against the peptide forms a specific immune complex with the protein.

Preferred epitopes encompassed by the antigenic peptide are regions that are located on the surface of the protein, e.g., hydrophilic regions. FIGS. 1L, 1M, 2J, 3U, 4I, 4J, 5E, 6F, and 7D are hydrophobicity plots of the proteins of the invention. These plots or similar analyses can be used to identify hydrophilic regions.

An immunogen typically is used to prepare antibodies by immunizing a suitable (i.e., immunocompetent) subject such as a rabbit, goat, mouse, or other mammal or vertebrate. An appropriate immunogenic preparation can contain, for example, recombinantly-expressed or chemically-synthesized polypeptide. The preparation can further include an adjuvant, such as Freund's complete or incomplete adjuvant, or a similar immunostimulatory agent.

Accordingly, another aspect of the invention pertains to antibodies directed against a polypeptide of the invention. The terms “antibody” and “antibody substance” as used interchangeably herein refer to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site which specifically binds an antigen, such as a polypeptide of the invention (e.g., an epitope of a polypeptide of the invention). A molecule which specifically binds to a given polypeptide of the invention is a molecule which binds the polypeptide, but does not substantially bind other molecules in a sample, e.g., a biological sample, which naturally contains the polypeptide. Examples of immunologically active portions of immunoglobulin molecules include F(ab) and F(ab′)₂ fragments which can be generated by treating the antibody with an enzyme such as pepsin. The invention provides polyclonal and monoclonal antibodies. The term “monoclonal antibody” or “monoclonal antibody composition”, as used herein, refers to a population of antibody molecules that contain only one species of an antigen binding site capable of immunoreacting with a particular epitope.

Polyclonal antibodies can be prepared as described above by immunizing a suitable subject with a polypeptide of the invention as an immunogen. Preferred polyclonal antibody compositions are ones that have been selected for antibodies directed against (i.e., which bind specifically with) one or more polypeptides of the invention. Particularly preferred polyclonal antibody preparations are ones that contain only antibodies directed against one or more polypeptides of the invention. Particularly preferred immunogen compositions are those that contain no other human proteins such as, for example, immunogen compositions made using a non-human host cell for recombinant expression of a polypeptide of the invention. In such a manner, the only human epitope or epitopes recognized by the resulting antibody compositions raised against this immunogen will be present as part of a polypeptide or polypeptides of the invention.

The antibody titer in the immunized subject can be monitored over time by standard techniques, such as with an enzyme linked immunosorbent assay (ELISA) using immobilized polypeptide. If desired, the antibody molecules can be harvested or isolated from the subject (e.g., from the blood or serum of the subject) and further purified by well-known techniques, such as protein A chromatography to obtain the IgG fraction. Alternatively, antibodies which bind specifically with a protein or polypeptide of the invention can be selected or purified (e.g., partially purified) using chromatographic methods, such as affinity chromatography. For example, a recombinantly expressed and purified (or partially purified) protein of the invention can be produced as described herein, and covalently or non-covalently coupled with a solid support such as, for example, a chromatography column. The column thus exhibits specific affinity for antibody substances which bind specifically with the protein of the invention, and these antibody substances can be purified from a sample containing antibody substances directed against a large number of different epitopes, thereby generating a substantially purified antibody substance composition, i.e., one that is substantially free of antibody substances which do not bind specifically with the protein. A substantially purified antibody composition, in this context, means an antibody sample that contains at most only 30% (by dry weight) of contaminating antibodies directed against epitopes other than those on the desired protein or polypeptide of the invention, preferably at most 20%, more preferably at most 10%, most preferably at most 5% (by dry weight of the sample is contaminating antibodies). A purified antibody composition means that at least 99% of the antibodies in the composition are directed against the desired protein or polypeptide of the invention.

At an appropriate time after immunization, e.g., when the specific antibody titers are highest, antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique originally described by Kohler and Milstein (1975) Nature 256:495-497, the human B cell hybridoma technique (Kozbor et al. (1983) Immunol. Today 4:72), the EBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96) or trioma techniques. The technology for producing hybridomas is well known (see generally Current Protocols in Immunology (1994) Coligan et al. (eds.) John Wiley & Sons, Inc., New York, N.Y.). Hybridoma cells producing a monoclonal antibody of the invention are detected by screening the hybridoma culture supernatants for antibodies that bind the polypeptide of interest, e.g., using a standard ELISA assay.

Alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal antibody directed against a polypeptide of the invention can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) with the polypeptide of interest. Kits for generating and screening phage display libraries are commercially available (e.g., the Pharmacia Recombinant Phage Antibody System, Catalog No. 27-9400-01; and the Stratagene SURFZAP™ Phage Display Kit, Catalog No. 240612). Additionally, examples of methods and reagents particularly amenable for use in generating and screening antibody display library can be found in, for example, U.S. Pat. No. 5,223,409; PCT Publication No. WO 92/18619; PCT Publication No. WO 91/17271; PCT Publication No. WO 92,20791; PCT Publication No. WO 92/15679; PCT Publication No. WO 93/01288; PCT Publication No. WO 92/01047; PCT Publication No. WO 92/09690; PCT Publication No. WO 90/02809; Fuchs et al. (1991) Bio/Technology 9:1370-1372; Hay et al. (1992) Hum. Antibod. Hybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffiths et al. (1993) EMBO J. 12:725-734.

Additionally, recombinant antibodies, such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, which can be made using standard recombinant DNA techniques, are within the scope of the invention. A chimeric antibody is a molecule in which different portions of the antibody amino acid sequence are derived from different animal species, such as those having a variable region derived from a murine monoclonal antibody and a constant region derived from a human immunoglobulin. (See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; and Boss et al., U.S. Pat. No. 4,816,397). Humanized antibodies are antibody molecules which are obtained from non-human species, which have one or more complementarity-determining regions (CDRs) derived from the non-human species, and which have a framework region derived from a human immunoglobulin molecule. (See, e.g., Queen, U.S. Pat. No. 5,585,089). Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example using methods described in PCT Publication No. WO 87/02671; European Patent Application 184,187; European Patent Application 171,496; European Patent Application 173,494; PCT Publication No. WO 86/01533; U.S. Pat. No. 4,816,567; European Patent Application 125,023; Better et al. (1988) Science 240:1041-1043; Liu et al. (1987) Proc. Natl. Acad. Sci. USA 84:3439-3443; Liu et al. (1987) J. Immunol. 139:3521-3526; Sun et al. (1987) Proc. Natl. Acad. Sci. USA 84:214-218; Nishimura et al. (1987) Canc. Res. 47:999-1005; Wood et al. (1985) Nature 314:446-449; and Shaw et al. (1988) J. Natl. Cancer Inst. 80:1553-1559); Morrison (1985) Science 229:1202-1207; Oi et al. (1986) Bio/Techniques 4:214; U.S. Pat. No. 5,225,539; Jones et al. (1986) Nature 321:552-525; Verhoeyan et al. (1988) Science 239:1534; and Beidler et al. (1988) J. Immunol. 141:4053-4060.

Completely human antibodies are particularly desirable for therapeutic treatment of human patients. Such antibodies can be produced, for example, using transgenic mice which are incapable of expressing endogenous immunoglobulin heavy and light chains genes, but which can express human heavy and light chain genes. The transgenic mice are immunized in the normal fashion with a selected antigen, e.g., all or a portion of a polypeptide of the invention. Monoclonal antibodies directed against the antigen can be obtained using conventional hybridoma technology. The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation. Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA and IgE antibodies. For an overview of this technology for producing human antibodies, see Lonberg and Huszar (1995, Int. Rev. Immunol. 13:65-93). For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, e.g., U.S. Pat. Nos. 5,625,126; 5,633,425; 5,569,825; 5,661,016; and 5,545,806. In addition, companies such as Abgenix, Inc. (Freemont, Calif.), can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above.

Completely human antibodies which recognize a selected epitope can be generated using a technique referred to as “guided selection.” In this approach a selected non-human monoclonal antibody, e.g., a murine antibody, is used to guide the selection of a completely human antibody recognizing the same epitope (Jespers et al. (1994) Bio/technology 12:899-903).

An antibody directed against a polypeptide of the invention (e.g., monoclonal antibody) can be used to isolate the polypeptide by standard techniques, such as affinity chromatography or immunoprecipitation. Moreover, such an antibody can be used to detect the protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the polypeptide. The antibodies can also be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or ³H.

Further, an antibody substance can be conjugated with a therapeutic moiety such as a cytotoxin, a therapeutic agent, or a radioactive metal ion. Cytotoxins and cytotoxic agents include any agent that is detrimental to cells. Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithrarnycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, and analogs or homologs of these compounds. Therapeutic agents include, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, and decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine {BSNU}, lomustine {CCNU}, cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin {formerly daunomycin} and doxorubicin), antibiotics (e.g., dactinomycin {formerly actinomycin}, bleomycin, mithramycin, and anthramycin {AMC}), and anti-mitotic agents (e.g., vincristine and vinblastine).

The conjugates of the invention can be used to modify a biological response; the drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety can be a protein or polypeptide which exhibits a desired biological activity. Such proteins include, for example, toxins such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; proteins such as tumor necrosis factor, alpha-interferon, beta-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator; and biological response modifiers such as lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), and other growth factors.

Techniques for conjugating a therapeutic moiety with an antibody substance are well known (see, e.g., Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies and Cancer Therapy, Reisfeld et al., eds., pp. 243-256, Alan R. Liss, Inc., 1985; Hellstrom et al., “Antibodies For Drug Delivery”, in Controlled Drug Delivery, 2nd Ed., Robinson et al., eds., pp. 623-653, Marcel Dekker, Inc., 1987; Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies '84: Biological and Clinical Applications, Pinchera et al., eds., pp. 475-506, 1985; “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies for Cancer Detection and Therapy, Baldwin et al., eds., pp. 303-316, Academic Press, 1985; and Thorpe et al., “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”, Immunol. Rev. 62:119-58, 1982). Alternatively, an antibody can be conjugated with a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980.

Accordingly, in one aspect, the invention provides substantially purified antibodies or fragment thereof, and non-human antibodies or fragments thereof, which antibodies or fragments specifically bind with a polypeptide having an amino acid sequence which comprises a sequence selected from the group consisting of:

(i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(ii) the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221;

(iii) at least 15 amino acid residues of the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(iv) an amino acid sequence which is at least 95% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, wherein the percent identity is determined using the ALIGN program of the GCG software package with a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4; and

(v) an amino acid sequence which is encoded by a nucleic acid molecule which hybridizes with a nucleic acid having a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions of hybridization of 6×SSC (standard saline citrate) at 45° C. and washing in 0.2×SSC, 0.1% SDS at 65° C.

In another aspect, the invention provides non-human antibodies or fragments thereof, which antibodies or fragments specifically bind with a polypeptide having an amino acid sequence which comprises a sequence selected from the group consisting of:

(i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(ii) the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221;

(iii) at least 15 amino acid residues of the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(iv) an amino acid sequence which is at least 95% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, wherein the percent identity is determined using the ALIGN program of the GCG software package with a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4; and

(v) an amino acid sequence which is encoded by a nucleic acid molecule which hybridizes with a nucleic acid having a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions of hybridization of 6×SSC (standard saline citrate) at 45° C. and washing in 0.2×SSC, 0.1% SDS at 65° C. Such non-human antibodies can be goat, mouse, sheep, horse, chicken, rabbit, or rat antibodies. Alternatively, the non-human antibodies of the invention can be chimeric and/or humanized antibodies. In addition, the non-human antibodies of the invention can be polyclonal antibodies or monoclonal antibodies.

In still a further aspect, the invention provides monoclonal antibodies or fragments thereof, which antibodies or fragments specifically bind with a polypeptide having an amino acid sequence which comprises a sequence selected from the group consisting of:

(i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(ii) the amino acid sequence encoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221;

(iii) at least 15 amino acid residues of the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(iv) an amino acid sequence which is at least 95% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, wherein the percent identity is determined using the ALIGN program of the GCG software package with a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4; and

(v) an amino acid sequence which is encoded by a nucleic acid molecule which hybridizes with a nucleic acid having a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions of hybridization of 6×SSC (standard saline citrate) at 45° C. and washing in 0.2×SSC, 0.1% SDS at 65° C. The monoclonal antibodies can be human, humanized, chimeric and/or non-human antibodies.

The substantially purified antibodies or fragments thereof can specifically bind with a signal peptide, a secreted sequence, an extracellular domain, a transmembrane or a cytoplasmic domain cytoplasmic membrane of a polypeptide of the invention. In a particularly preferred embodiment, the substantially purified antibodies or fragments thereof, the non-human antibodies or fragments thereof, and/or the monoclonal antibodies or fragments thereof, of the invention specifically bind with a secreted sequence or with an extracellular domain of one of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. Preferably, the extracellular domain with which the antibody substance binds has an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 14, 22, 30, 37, 49, 50, and 56-58.

Any of the antibody substances of the invention can be conjugated with a therapeutic moiety or to a detectable substance. Non-limiting examples of detectable substances that can be conjugated with the antibody substances of the invention include an enzyme, a prosthetic group, a fluorescent material (i.e., a fluorophore), a luminescent material, a bioluminescent material, and a radioactive material (e.g., a radionuclide or a substituent comprising a radionuclide).

The invention also provides a kit containing an antibody substance of the invention conjugated with a detectable substance, and instructions for use. Still another aspect of the invention is a pharmaceutical composition comprising an antibody substance of the invention and a pharmaceutically acceptable carrier. In preferred embodiments, the pharmaceutical composition contains an antibody substance of the invention, a therapeutic moiety (preferably conjugated with the antibody substance), and a pharmaceutically acceptable carrier.

Still another aspect of the invention is a method of making an antibody that specifically recognizes one of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. This method comprises immunizing a vertebrate (e.g., a mammal such as a rabbit, goat, or pig) with a polypeptide. The polypeptide used as an immunogen has an amino acid sequence that comprises a sequence selected from the group consisting of:

(i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(ii) the amino acid sequence encoded by a cDNA of a clone deposited as one of ATTC® 207219, 207184, 207228, 207185, 207220, and 207221;

(iii) at least 15 amino acid residues of the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74;

(iv) an amino acid sequence which is at least 95% identical to the amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, wherein the percent identity is determined using the ALIGN program of the GCG software package with a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4; and

(v) an amino acid sequence which is encoded by a nucleic acid molecule which hybridizes with a nucleic acid having a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions of hybridization of 6×SSC (standard saline citrate) at 45° C. and washing in 0.2×SSC, 0.1% SDS at 65° C.

After immunization, a sample is collected from the vertebrate that contains an antibody that specifically recognizes the polypeptide with which the vertebrate was immunized. Preferably, the polypeptide is recombinantly produced using a non-human host cell. Optionally, an antibody substance can be further purified from the sample using techniques well known to those of skill in the art. The method can further comprise making a monoclonal antibody-producing cell from a cell of the vertebrate. Optionally, antibodies can be collected from the antibody-producing cell.

III. Recombinant Expression Vectors and Host Cells

Another aspect of the invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding a polypeptide of the invention (or a portion thereof). As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors, expression vectors, are capable of directing the expression of genes to which they are operably linked. In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids (vectors). However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses; adenoviruses and adeno-associated viruses), which serve equivalent functions.

The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell. This means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operably linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, “operably linked” is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term “regulatory sequence” is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those which direct constitutive expression of a nucleotide sequence in many types of host cell and those which direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, and the like. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein.

The recombinant expression vectors of the invention can be designed for expression of a polypeptide of the invention in prokaryotic (e.g., E. coli) or eukaryotic cells (e.g., insect cells (using baculovirus expression vectors), yeast cells or mammalian cells). Suitable host cells are discussed further in Goeddel, supra. Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

Expression of proteins in prokaryotes is most often carried out in E. coli a with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: 1) to increase expression of recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson (1988) Gene 67:3140), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.

Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amann et al., (1988) Gene 69:301-315) and pET 11d (Studier et al., Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990) 60-89). Target gene expression from the pTrc vector relies on host RNA polymerase transcription from a hybrid trp-lac fusion promoter. Target gene expression from the pET 11d vector relies on transcription from a T7 gn10-lac fusion promoter mediated by a co-expressed viral RNA polymerase (T7 gn1). This viral polymerase is supplied by host strains BL21 (DE3) or HMS174(DE3) from a resident lambda prophage harboring a T7 gn1 gene under the transcriptional control of the lacUV 5 promoter.

One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein (Gottesman, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990) 119-128). Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (Wada et al. (1992) Nucleic Acids Res. 20:2111-2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard DNA synthesis techniques.

In another embodiment, the expression vector is a yeast expression vector. Examples of vectors for expression in yeast S. cerevisiae include pYepSec1 (Baldari et al. (1987) EMBO J. 6:229-234), pMFa (Kuran and Herskowitz, (1982) Cell 30:933-943), pJRY88 (Schultz et al. (1987) Gene 54:113-123), pYES2 (Invitrogen Corporation, San Diego, Calif.), and pPicZ (Invitrogen Corp, San Diego, Calif.).

Alternatively, the expression vector is a baculovirus expression vector. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf 9 cells) include the pAc series (Smith et al. (1983) Mol. Cell Biol. 3:2156-2165) and the pVL series (Lucklow and Summers (1989) Virology 170:31-39).

In yet another embodiment, a nucleic acid of the invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed (1987) Nature 329:840) and pMT2PC (Kaufman et al. (1987) EMBO J. 6:187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, Adenovirus 2, cytomegalovirus and Simian Virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see chapters 16 and 17 of Sambrook et al., supra.

In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art. Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert et al. (1987) Genes Dev. 1:268-277), lymphoid-specific promoters (Calame and Eaton (1988) Adv. Immunol. 43:235-275), in particular promoters of T cell receptors (Winoto and Baltimore (1989) EMBO J. 8:729-733) and immunoglobulins (Banerji et al. (1983) Cell 33:729-740; Queen and Baltimore (1983) Cell 33:741-748), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle (1989) Proc. Natl. Acad. Sci. USA 86:5473-5477), pancreas-specific promoters (Edlund et al. (1985) Science 230:912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, for example the murine hox promoters (Kessel and Gruss (1990) Science 249:374-379) and the α-fetoprotein promoter (Campes and Tilghman (1989) Genes Dev. 3:537-546).

The invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operably linked to a regulatory sequence in a manner which allows for expression (by transcription of the DNA molecule) of an RNA molecule which is antisense to the mRNA encoding a polypeptide of the invention. Regulatory sequences operably linked to a nucleic acid cloned in the antisense orientation can be chosen which direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen which direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid, or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion of the regulation of gene expression using antisense genes see Weintraub et al. (Reviews—Trends in Genetics, Vol. 1(1)1986).

Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

A host cell can be any prokaryotic (e.g., E. coli) or eukaryotic cell (e.g., insect cells, yeast or mammalian cells).

Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (supra), and other laboratory manuals.

For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., for resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Preferred selectable markers include those which confer resistance to drugs, such as G418, hygromycin and methotrexate. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die).

In another embodiment, the expression characteristics of an endogenous nucleic acid within a cell, cell line, or microorganism (e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 nucleic acid, as described herein) can be modified by inserting a heterologous DNA regulatory element (i.e., one that is heterologous with respect to the endogenous gene) into the genome of the cell, stable cell line, or cloned microorganism. The inserted regulatory element can be operatively linked with the endogenous gene (e.g., TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296) and thereby control, modulate, or activate the endogenous gene. For example, an endogenous TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene which is normally “transcriptionally silent” (i.e., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene which is normally not expressed, or is normally expressed only at only a very low level) can be activated by inserting a regulatory element which is capable of promoting expression of the gene in the cell, cell line, or microorganism. Alternatively, a transcriptionally silent, endogenous TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene can be activated by inserting a promiscuous regulatory element that works across cell types.

A heterologous regulatory element can be inserted into a stable cell line or cloned microorganism such that it is operatively linked with and activates expression of an endogenous TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene, using techniques, such as targeted homologous recombination, which are well known to those of skill in the art (described e.g., in Chappel, U.S. Pat. No. 5,272,071; PCT publication No. WO 91/06667, published May 16, 1991).

A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce a polypeptide of the invention. Accordingly, the invention further provides methods for producing a polypeptide of the invention using the host cells of the invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding a polypeptide of the invention has been introduced) in a suitable medium such that the polypeptide is produced. In another embodiment, the method further comprises isolating the polypeptide from the medium or the host cell.

The host cells of the invention can also be used to produce non-human transgenic animals. For example, in one embodiment, a host cell of the invention is a fertilized oocyte or an embryonic stem cell into which a sequences encoding a polypeptide of the invention have been introduced. Such host cells can then be used to create non-human transgenic animals in which exogenous sequences encoding a polypeptide of the invention have been introduced into their genome or homologous recombinant animals in which endogenous encoding a polypeptide of the invention sequences have been altered. Such animals are useful for studying the function and/or activity of the polypeptide and for identifying and/or evaluating modulators of polypeptide activity. As used herein, a “transgenic animal” is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more of the cells of the animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, etc. A transgene is exogenous DNA which is integrated into the genome of a cell from which a transgenic animal develops and which remains in the genome of the mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues of the transgenic animal. As used herein, an “homologous recombinant animal” is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal, e.g., an embryonic cell of the animal, prior to development of the animal.

A transgenic animal of the invention can be created by introducing nucleic acid encoding a polypeptide of the invention (or a homologue thereof) into the male pronuclei of a fertilized oocyte, e.g., by microinjection, retroviral infection, and allowing the oocyte to develop in a pseudopregnant female foster animal. Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression of the transgene. A tissue-specific regulatory sequence(s) can be operably linked to the transgene to direct expression of the polypeptide of the invention to particular cells. Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat. Nos. 4,736,866 and 4,870,009, 4,873,191, in Hogan, Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986, and in Wakayama et al., 1999, Proc. Natl. Acad. Sci. USA 96:14984-14989. Similar methods can be used to produce other transgenic animals. A transgenic founder animal can be identified based upon the presence of the transgene in its genome and/or expression of mRNA encoding the transgene in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying the transgene can further be bred to other transgenic animals carrying other transgenes.

To create a homologous recombinant animal, a vector is prepared which contains at least a portion of a gene encoding a polypeptide of the invention into which a deletion, addition or substitution has been introduced to thereby alter, e.g., functionally disrupt, the gene. In a preferred embodiment, the vector is designed such that, upon homologous recombination, the endogenous gene is functionally disrupted (i.e., no longer encodes a functional protein; also referred to as a “knock out” vector). Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous protein). In the homologous recombination vector, the altered portion of the gene is flanked at its 5′ and 3′ ends by additional nucleic acid of the gene to allow for homologous recombination to occur between the exogenous gene carried by the vector and an endogenous gene in an embryonic stem cell. The additional flanking nucleic acid sequences are of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA (both at the 5′ and 3′ ends) are included in the vector (see, e.g., Thomas and Capecchi (1987) Cell 51:503 for a description of homologous recombination vectors). The vector is introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced gene has homologously recombined with the endogenous gene are selected (see, e.g., Li et al. (1992) Cell 69:915). The selected cells are then injected into a blastocyst of an animal (e.g., a mouse) to form aggregation chimeras (see, e.g., Bradley in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, Robertson, ed. (IRL, Oxford, 1987) pp. 113-152). A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term. Progeny harboring the homologously recombined DNA in their germ cells can be used to breed animals in which all cells of the animal contain the homologously recombined DNA by germline transmission of the transgene. Methods for constructing homologous recombination vectors and homologous recombinant animals are described further in Bradley (1991) Current Opinion in Bio/Technology 2:823-829 and in PCT Publication Nos. WO 90/11354, WO 91/01140, WO 92/0968, and WO 93/04169.

In another embodiment, transgenic non-human animals can be produced which contain selected systems which allow for regulated expression of the transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage P1. For a description of the cre/loxP recombinase system, see, e.g., Lakso et al. (1992) Proc. Natl. Acad. Sci. USA 89:6232-6236. Another example of a recombinase system is the FLP recombinase system of Saccharomyces cerevisiae (O'Gorman et al. (1991) Science 251:1351-1355. If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein are required. Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.

Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut et al. (1997) Nature 385:810-813 and PCT Publication Nos. WO 97/07668 and WO 97/07669.

IV. Pharmaceutical Compositions

The nucleic acid molecules, polypeptides, and antibodies (also referred to herein as “active compounds”) of the invention can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

The invention includes methods for preparing pharmaceutical compositions for modulating the expression or activity of a polypeptide or nucleic acid of the invention. Such methods comprise formulating a pharmaceutically acceptable carrier with an agent which modulates expression or activity of a polypeptide or nucleic acid of the invention. Such compositions can further include additional active agents. Thus, the invention further includes methods for preparing a pharmaceutical composition by formulating a pharmaceutically acceptable carrier with an agent which modulates expression or activity of a polypeptide or nucleic acid of the invention and one or more additional active compounds.

The agent which modulates expression or activity can, for example, be a small molecule other than a nucleic acid, polypeptide, or antibody of the invention. For example, such small molecules include peptides, peptidomimetics, amino acids, amino acid analogs, polynucleotides, polynucleotide analogs, nucleotides, nucleotide analogs, organic or inorganic compounds (i.e., including heteroorganic and organometallic compounds) having a molecular weight less than about 10,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 5,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 1,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 500 grams per mole, and salts, esters, and other pharmaceutically acceptable forms of such compounds.

It is understood that appropriate doses of small molecule agents and protein or polypeptide agents depends upon a number of factors within the ken of the ordinarily skilled physician, veterinarian, or researcher. The dose(s) of these agents will vary, for example, depending upon the identity, size, and condition of the subject or sample being treated, further depending upon the route by which the composition is to be administered, if applicable, and the effect which the practitioner desires the agent to have upon the nucleic acid or polypeptide of the invention. Exemplary doses of a small molecule include milligram or microgram amounts per kilogram of subject or sample weight (e.g., about 1 microgram per kilogram to about 500 milligrams per kilogram, about 100 micrograms per kilogram to about 5 milligrams per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram). Exemplary doses of a protein or polypeptide include gram, milligram or microgram amounts per kilogram of subject or sample weight (e.g., about 1 microgram per kilogram to about 5 grams per kilogram, about 100 micrograms per kilogram to about 500 milligrams per kilogram, or about 1 milligram per kilogram to about 50 milligrams per kilogram). It is furthermore understood that appropriate doses of one of these agents depend upon the potency of the agent with respect to the expression or activity to be modulated. Such appropriate doses can be determined using the assays described herein. When one or more of these agents is to be administered to an animal (e.g., a human) in order to modulate expression or activity of a polypeptide or nucleic acid of the invention, a physician, veterinarian, or researcher can, for example, prescribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained. In addition, it is understood that the specific dose level for any particular animal subject will depend upon a variety of factors including the activity of the specific agent employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, any drug combination, and the degree of expression or activity to be modulated.

A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediamine-tetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampules, disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL (BASF; Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound (e.g., a polypeptide or antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium, and then incorporating the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed.

Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches, and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressurized container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes which can be targeted to bind with virus-infected cells using a monoclonal antibody which binds specifically with a viral antigen) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

For antibodies, the preferred dosage is 0.1 mg/kg to 100 mg/kg of body weight (generally 10 mg/kg to 20 mg/kg). If the antibody is to act in the brain, a dosage of 50 mg/kg to 100 mg/kg is usually appropriate. Generally, partially human antibodies and fully human antibodies have a longer half-life within the human body than other antibodies. Accordingly, lower dosages and less frequent administration is often possible. Modifications such as lipidation can be used to stabilize antibodies and to enhance uptake and tissue penetration (e.g., into the brain). A method for lipidation of antibodies is described by Cruikshank et al. ((1997) J. Acquired Immune Deficiency Syndromes and Human Retrovirology 14:193).

The nucleic acid molecules of the invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (U.S. Pat. No. 5,328,470), or by stereotactic injection (see, e.g., Chen et al. (1994) Proc. Natl. Acad. Sci. USA 91:3054-3057). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g. retroviral vectors, the pharmaceutical preparation can include one or more cells which produce the gene delivery system.

The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

V. Uses and Methods of the Invention

The nucleic acid molecules, proteins, protein homologs, and antibodies described herein can be used in one or more of the following methods: a) screening assays; b) detection assays (e.g., chromosomal mapping, tissue typing, forensic biology); c) predictive medicine (e.g., diagnostic assays, prognostic assays, monitoring clinical trials, and pharmacogenomics); and d) methods of treatment (e.g., therapeutic and prophylactic). For example, polypeptides of the invention can to used for all of the purposes identified herein in portions of the disclosure relating to individual types of protein of the invention (e.g., TANGO 202 proteins, TANGO 234 proteins, TANGO 265 proteins, TANGO 273 proteins, TANGO 286 proteins, TANGO 294 proteins, and INTERCEPT 296 proteins). Polypeptides of the invention can also be used to modulate cellular proliferation, cellular differentiation, cellular adhesion, or some combination of these. The isolated nucleic acid molecules of the invention can be used to express proteins (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect mRNA (e.g., in a biological sample) or a genetic lesion, and to modulate activity of a polypeptide of the invention. In addition, the polypeptides of the invention can be used to screen drugs or compounds which modulate activity or expression of a polypeptide of the invention as well as to treat disorders characterized by insufficient or excessive production of a protein of the invention or production of a form of a protein of the invention which has decreased or aberrant activity compared to the wild type protein. In addition, the antibodies of the invention can be used to detect and isolate a protein of the and modulate activity of a protein of the invention.

This invention further pertains to novel agents identified by the above-described screening assays and uses thereof for treatments as described herein.

A. Screening Assays

The invention provides a method (also referred to herein as a “screening assay”) for identifying modulators, i.e., candidate or test compounds or agents (e.g., peptides, peptidomimetics, small molecules or other drugs) which bind to polypeptide of the invention or have a stimulatory or inhibitory effect on, for example, expression or activity of a polypeptide of the invention.

In one embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity of the membrane-bound form of a polypeptide of the invention or biologically active portion thereof. The test compounds of the present invention can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the “one-bead one-compound” library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds (Lam (1997) Anticancer Drug Des. 12:145).

Examples of methods for the synthesis of molecular libraries can be found in the art for example in: DeWitt et al. (1993) Proc. Natl. Acad. Sci. USA 90:6909; Erb et al. (1994) Proc. Natl. Acad. Sci. USA 91:11422; Zuckermann et al. (1994). J. Med. Chem. 37:2678; Cho et al. (1993) Science 261:1303; Carrell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2059; Carell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2061; and Gallop et al. (1994) J. Med. Chem. 37:1233.

Libraries of compounds can be presented in solution (e.g., Houghten (1992) Bio/Techniques 13:412-421), or on beads (Lam (1991) Nature 354:82-84), chips (Fodor (1993) Nature 364:555-556), bacteria (U.S. Pat. No. 5,223,409), spores (U.S. Pat. Nos. 5,571,698; 5,403,484; and 5,223,409), plasmids (Cull et al. (1992) Proc. Natl. Acad. Sci. USA 89:1865-1869) or phage (Scott and Smith (1990) Science 249:386-390; Devlin (1990) Science 249:404-406; Cwirla et al. (1990) Proc. Natl. Acad. Sci. USA 87:6378-6382; and Felici (1991) J. Mol. Biol. 222:301-310).

In one embodiment, an assay is a cell-based assay in which a cell which expresses a membrane-bound form of a polypeptide of the invention, or a biologically active portion thereof, on the cell surface is contacted with a test compound and the ability of the test compound to bind to the polypeptide determined. The cell, for example, can be a yeast cell or a cell of mammalian origin. Determining the ability of the test compound to bind to the polypeptide can be accomplished, for example, by coupling the test compound with a radioisotope or enzymatic label such that binding of the test compound to the polypeptide or biologically active portion thereof can be determined by detecting the labeled compound in a complex. For example, test compounds can be labeled with ¹²⁵I, ³⁵S, ¹⁴C, or ³H, either directly or indirectly, and the radioisotope detected by direct counting of radio-emission or by scintillation counting. Alternatively, test compounds can be enzymatically labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. In a preferred embodiment, the assay comprises contacting a cell which expresses a membrane-bound form of a polypeptide of the invention, or a biologically active portion thereof, on the cell surface with a known compound which binds the polypeptide to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with the polypeptide, wherein determining the ability of the test compound to interact with the polypeptide comprises determining the ability of the test compound to preferentially bind to the polypeptide or a biologically active portion thereof as compared to the known compound.

In another embodiment, the assay involves assessment of an activity characteristic of the polypeptide, wherein binding of the test compound with the polypeptide or a biologically active portion thereof alters (i.e., increases or decreases) the activity of the polypeptide.

In another embodiment, an assay is a cell-based assay comprising contacting a cell expressing a membrane-bound form of a polypeptide of the invention, or a biologically active portion thereof, on the cell surface with a test compound and determining the ability of the test compound to modulate (e.g., stimulate or inhibit) the activity of the polypeptide or biologically active portion thereof. Determining the ability of the test compound to modulate the activity of the polypeptide or a biologically active portion thereof can be accomplished, for example, by determining the ability of the polypeptide to bind to or interact with a target molecule or to transport molecules across the cytoplasmic membrane.

Determining the ability of a polypeptide of the invention to bind to or interact with a target molecule can be accomplished by one of the methods described above for determining direct binding. As used herein, a “target molecule” is a molecule with which a selected polypeptide (e.g., a polypeptide of the invention binds or interacts with in nature, for example, a molecule on the surface of a cell which expresses the selected protein, a molecule on the surface of a second cell, a molecule in the extracellular milieu, a molecule associated with the internal surface of a cell membrane or a cytoplasmic molecule. A target molecule can be a polypeptide of the invention or some other polypeptide or protein. For example, a target molecule can be a component of a signal transduction pathway which facilitates transduction of an extracellular signal (e.g., a signal generated by binding of a compound to a polypeptide of the invention) through the cell membrane and into the cell or a second intercellular protein which has catalytic activity or a protein which facilitates the association of downstream signaling molecules with a polypeptide of the invention. Determining the ability of a polypeptide of the invention to bind to or interact with a target molecule can be accomplished by determining the activity of the target molecule. For example, the activity of the target molecule can be determined by detecting induction of a cellular second messenger of the target (e.g., an mRNA, intracellular Ca²⁺, diacylglycerol, IP3, and the like), detecting catalytic/enzymatic activity of the target on an appropriate substrate, detecting the induction of a reporter gene (e.g., a regulatory element that is responsive to a polypeptide of the invention operably linked to a nucleic acid encoding a detectable marker, e.g. luciferase), or detecting a cellular response, for example, cellular differentiation, or cell proliferation.

In yet another embodiment, an assay of the present invention is a cell-free assay comprising contacting a polypeptide of the invention or biologically active portion thereof with a test compound and determining the ability of the test compound to bind to the polypeptide or biologically active portion thereof. Binding of the test compound to the polypeptide can be determined either directly or indirectly as described above. In a preferred embodiment, the assay includes contacting the polypeptide of the invention or biologically active portion thereof with a known compound which binds the polypeptide to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with the polypeptide, wherein determining the ability of the test compound to interact with the polypeptide comprises determining the ability of the test compound to preferentially bind to the polypeptide or biologically active portion thereof as compared to the known compound.

In another embodiment, an assay is a cell-free assay comprising contacting a polypeptide of the invention or biologically active portion thereof with a test compound and determining the ability of the test compound to modulate (e.g., stimulate or inhibit) the activity of the polypeptide or biologically active portion thereof. Determining the ability of the test compound to modulate the activity of the polypeptide can be accomplished, for example, by determining the ability of the polypeptide to bind to a target molecule by one of the methods described above for determining direct binding. In an alternative embodiment, determining the ability of the test compound to modulate the activity of the polypeptide can be accomplished by determining the ability of the polypeptide of the invention to further modulate the target molecule. For example, the catalytic activity, the enzymatic activity, or both, of the target molecule on an appropriate substrate can be determined as previously described.

In yet another embodiment, the cell-free assay comprises contacting a polypeptide of the invention or biologically active portion thereof with a known compound which binds the polypeptide to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with the polypeptide, wherein determining the ability of the test compound to interact with the polypeptide comprises determining the ability of the polypeptide to preferentially bind to or modulate the activity of a target molecule.

The cell-free assays of the present invention are amenable to use of both a soluble form or the membrane-bound form of a polypeptide of the invention. In the case of cell-free assays comprising the membrane-bound form of the polypeptide, it can be desirable to utilize a solubilizing agent such that the membrane-bound form of the polypeptide is maintained in solution. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-octylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton X-100, Triton X-114, Thesit, Isotridecypoly(ethylene glycol ether)n, 3-[(3-cholamidopropyl)dimethylamminio]-1-propane sulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylamminio]-2-hydroxy-1-propane sulfonate (CHAPSO), or N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate.

In one or more embodiments of the above assay methods of the present invention, it can be desirable to immobilize either the polypeptide of the invention or its target molecule to facilitate separation of complexed from non-complexed forms of one or both of the proteins, as well as to accommodate automation of the assay. Binding of a test compound to the polypeptide, or interaction of the polypeptide with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided which adds a domain that allows one or both of the proteins to be bound to a matrix. For example, glutathione-S-transferase fusion proteins or glutathione-S-transferase fusion proteins can be adsorbed onto glutathione SEPHAROSE™ beads (Sigma Chemical; St. Louis, Mo.) or glutathione derivatized microtiter plates, which are then combined with the test compound or the test compound and either the non-adsorbed target protein or A polypeptide of the invention, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtiter plate wells are washed to remove any unbound components and complex formation is measured either directly or indirectly, for example, as described above. Alternatively, the complexes can be dissociated from the matrix, and the level of binding or activity of the polypeptide of the invention can be determined using standard techniques.

Other techniques for immobilizing proteins on matrices can also be used in the screening assays of the invention. For example, either the polypeptide of the invention or its target molecule can be immobilized uitilizing conjugation of biotin and streptavidin. Biotinylated polypeptide of the invention or target molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well known in the art (e.g., biotinylation kit, Pierce Chemicals; Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with the polypeptide of the invention or target molecules but which do not interfere with binding of the polypeptide of the invention to its target molecule can be derivatized to the wells of the plate, and unbound target or polypeptide of the invention trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the polypeptide of the invention or target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the polypeptide of the invention or target molecule.

In another embodiment, modulators of expression of a polypeptide of the invention are identified in a method in which a cell is contacted with a candidate compound and the expression of the selected mRNA or protein (i.e., the mRNA or protein corresponding to a polypeptide or nucleic acid of the invention) in the cell is determined. The level of expression of the selected mRNA or protein in the presence of the candidate compound is compared to the level of expression of the selected mRNA or protein in the absence of the candidate compound. The candidate compound can then be identified as a modulator of expression of the polypeptide of the invention based on this comparison. For example, when expression of the selected mRNA or protein is greater (i.e., statistically significantly greater) in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of the selected mRNA or protein expression. Alternatively, when expression of the selected mRNA or protein is less (i.e., statistically significantly less) in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of the selected mRNA or protein expression. The level of the selected mRNA or protein expression in the cells can be determined by methods described herein.

In yet another aspect of the invention, a polypeptide of the inventions can be used as “bait proteins” in a two-hybrid assay or three hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993) Cell 72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054; Bartel et al. (1993) Bio/Techniques 14:920-924; Iwabuchi et al. (1993) Oncogene 8:1693-1696; and PCT Publication No. WO 94/10300), to identify other proteins, which bind to or interact with the polypeptide of the invention and modulate activity of the polypeptide of the invention. Such binding proteins are also likely to be involved in the propagation of signals by the polypeptide of the inventions as, for example, upstream or downstream elements of a signaling pathway involving the polypeptide of the invention.

This invention further pertains to novel agents identified by the above-described screening assays and uses thereof for treatments as described herein.

B. Detection Assays

Portions or fragments of the cDNA sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. For example, these sequences can be used to: (i) map their respective genes on a chromosome and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. These applications are described in the subsections below.

1. Chromosome Mapping

Once the sequence (or a portion of the sequence) of a gene has been isolated, this sequence can be used to map the location of the gene on a chromosome. Accordingly, nucleic acid molecules described herein or fragments thereof, can be used to map the location of the corresponding genes on a chromosome. The mapping of the sequences to chromosomes is an important first step in correlating these sequences with genes associated with disease.

Briefly, genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 base pairs in length) from the sequence of a gene of the invention. Computer analysis of the sequence of a gene of the invention can be used to rapidly select primers that do not span more than one exon in the genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the gene sequences will yield an amplified fragment. For a review of this technique, see D'Eustachio et al. ((1983) Science 220:919-924).

PCR mapping of somatic cell hybrids is a rapid procedure for assigning a particular sequence to a particular chromosome. Three or more sequences can be assigned per day using a single thermal cycler. Using the nucleic acid sequences of the invention to design oligonucleotide primers, sub-localization can be achieved with panels of fragments from specific chromosomes. Other mapping strategies which can similarly be used to map a gene to its chromosome include in situ hybridization (described in Fan et al. (1990) Proc. Natl. Acad. Sci. USA 87:6223-27), pre-screening with labeled flow-sorted chromosomes, and pre-selection by hybridization to chromosome specific cDNA libraries. Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. For a review of this technique, see Verma et al. (Human Chromosomes: A Manual of Basic Techniques (Pergamon Press, New York, 1988)).

Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to non-coding regions of the genes actually are preferred for mapping purposes. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping.

Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data. (Such data are found, for example, in V. McKusick, Mendelian Inheritance in Man, available on-line through Johns Hopkins University Welch Medical Library). The relationship between genes and disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, e.g., Egeland et al. (1987) Nature 325:783-787.

Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with a gene of the invention can be determined. If a mutation is observed in some or all of the affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent of the particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymorphisms.

Furthermore, the nucleic acid sequences disclosed herein can be used to perform searches against “mapping databases”, e.g., BLAST-type search, such that the chromosome position of the gene is identified by sequence homology or identity with known sequence fragments which have been mapped to chromosomes.

In the instant case, the human gene for TANGO 265 is located on chromosome 1 between markers D1S305 and D1S2635, and the human gene for TANGO 273 is located on chromosome 7 between markers D7S2467 and D7S2552.

In the instant case, the human gene for TANGO 286 exhibits significant amino acid homology with a region of the human chromosome region 22q12-13 genomic nucleotide sequence having GenBank Accession number AL021937. Alignment of a 45 kilobase nucleotide sequence encoding TANGO 286 with AL021937, however, indicated the presence in TANGO 286 of exons which differ from those disclosed in L021937 (pam120.mat scoring matrix; gap penalties −12/−4). This region of chromosome 22 comprises an immunoglobulin lambda chain C (IGLC) pseudogene, the Ret finger protein-like 3 (RFPL3) and Ret finger protein-like 3 antisense (RFPL3S) genes, a gene encoding a novel immunoglobulin lambda chain V family protein, a novel gene encoding a protein similar both to mouse RGDS protein (RALGDS, RALGEF, guanine nucleotide dissociation stimulator A) and to rabbit oncogene RSC, a novel gene encoding the human orthologue of worm F16A11.2 protein, a novel gene encoding a protein similar both to BPI and to rabbit liposaccharide-binding protein, and a 5′-portion of a novel gene. This region also comprises various ESTs, STSs, GSSs, genomic marker D22S1175, a ca repeat polymorphism and putative CpG islands.

A polypeptide and fragments and sequences thereof and antibodies which bind specifically with such polypeptides/fragments can be used to map the location of the gene encoding the polypeptide on a chromosome. This mapping can be performed by specifically detecting the presence of the polypeptide/fragments in members of a panel of somatic cell hybrids between cells obtained from a first species of animal from which the protein originates and cells obtained from a second species of animal, determining which somatic cell hybrid(s) expresses the polypeptide, and noting the chromosome(s) of the first species of animal that it contains. For examples of this technique (see Pajunen et al., 1988, Cytogenet. Cell Genet. 47:37-41 and Van Keuren et al., 1986, Hum. Genet. 74:34-40). Alternatively, the presence of the polypeptide in the somatic cell hybrids can be determined by assaying an activity or property of the polypeptide (e.g., enzymatic activity, as described in Bordelon-Riser et al., 1979, Som. Cell Genet. 5:597-613 and Owerbach et al., 1978, Proc. Natl. Acad. Sci. USA 75:5640-5644).

In the instant case, the human gene for TANGO 234 protein indicated that the gene is located at chromosomal location h12p13. Flanking chromosomal markers include WI-6980 and GATA8A09.43. Nearby human loci include IBD2 (inflammatory bowel disease 2), FPF (familial periodic fever), and HPDR2 (hypophosphatemia vitamin D resistant rickets 2). Nearby genes are KLRC (killer cell receptor cluster), DRPLA (dentatorubro-pallidoluysian atrophy), GAPD (glyceraldehyde-3-phosphate)dehydrogenase, and PXR1 (peroxisome receptor 1). This region is syntenic to mouse chromosome mo6. Murine chromosomal mapping indicated that the murine orthologue is located near the scr (scruffy) locus. Nearby mouse genes include drpla (dentatorubral phillidoluysian atrophy), prp (proline rich protein), and kap (kidney androgen regulated protein).

2. Tissue Typing

The nucleic acid sequences of the present invention can also be used to identify individuals from minute biological samples. The United States military, for example, is considering the use of restriction fragment length polymorphism (RFLP) for identification of its personnel. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identification. This method does not suffer from the current limitations of “Dog Tags” which can be lost, switched, or stolen, making positive identification difficult. The sequences of the present invention are useful as additional DNA markers for RFLP (described in U.S. Pat. No. 5,272,057).

Furthermore, the sequences of the present invention can be used to provide an alternative technique which determines the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the nucleic acid sequences described herein can be used to prepare two PCR primers from the 5′ and 3′ ends of the sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it.

Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences. The sequences of the present invention can be used to obtain such identification sequences from individuals and from tissue. The nucleic acid sequences of the invention uniquely represent portions of the human genome. Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the non-coding regions. It is estimated that allelic variation between individual humans occurs with a frequency of about once per each 500 bases. Each of the sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification purposes. Because greater numbers of polymorphisms occur in the non-coding regions, fewer sequences are necessary to differentiate individuals. The non-coding sequences of any of SEQ ID NOs: 1, 9, 17, 25, 33, 45, and 53 can comfortably provide positive individual identification with a panel of perhaps 10 to 1,000 primers which each yield a non-coding amplified sequence of 100 bases. If predicted coding sequences, such as those in any of SEQ ID NOs: 2, 10, 18, 26, 34, 46, and 54 are used, a more appropriate number of primers for positive individual identification would be 500-2,000.

If a panel of reagents from the nucleic acid sequences described herein is used to generate a unique identification database for an individual, those same reagents can later be used to identify tissue from that individual. Using the unique identification database, positive identification of the individual, living or dead, can be made from extremely small tissue samples.

3. Use of Partial Gene Sequences in Forensic Biology

DNA-based identification techniques can also be used in forensic biology. Forensic biology is a scientific field employing genetic typing of biological evidence found at a crime scene as a means for positively identifying, for example, a perpetrator of a crime. To make such an identification, PCR technology can be used to amplify DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, or semen found at a crime scene. The amplified sequence can then be compared to a standard, thereby allowing identification of the origin of the biological sample.

The sequences of the present invention can be used to provide polynucleotide reagents, e.g., PCR primers, targeted to specific loci in the human genome, which can enhance the reliability of DNA-based forensic identifications by, for example, providing another “identification marker” (i.e., another DNA sequence that is unique to a particular individual). As mentioned above, actual base sequence information can be used for identification as an accurate alternative to patterns formed by restriction enzyme generated fragments. Sequences targeted to non-coding regions are particularly appropriate for this use as greater numbers of polymorphisms occur in the non-coding regions, making it easier to differentiate individuals using this technique. Examples of polynucleotide reagents include the nucleic acid sequences of the invention or portions thereof, e.g., fragments derived from non-coding regions having a length of at least 20 or 30 bases.

The nucleic acid sequences described herein can further be used to provide polynucleotide reagents, e.g., labeled or labelable probes which can be used in, for example, an in situ hybridization technique, to identify a specific tissue, e.g., brain tissue. This can be very useful in cases where a forensic pathologist is presented with a tissue of unknown origin. Panels of such probes can be used to identify tissue by species and/or by organ type.

C. Predictive Medicine

The present invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenomics, and monitoring clinical trials are used for prognostic (predictive) purposes to thereby treat an individual prophylactically. Accordingly, one aspect of the present invention relates to diagnostic assays for determining expression of a polypeptide or nucleic acid of the invention and/or activity of a polypeptide of the invention (e.g., expression or activity of one of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 genes or proteins), in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or disorder, or is at risk of developing a disorder, associated with aberrant expression or activity of a polypeptide of the invention. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing a disorder associated with aberrant expression or activity of a polypeptide of the invention. For example, mutations in a gene of the invention can be assayed in a biological sample. Such assays can be used for prognostic or predictive purpose to thereby prophylactically treat an individual prior to the onset of a disorder characterized by or associated with aberrant expression or activity of a polypeptide of the invention.

As an alternative to making determinations based on the absolute expression level of a selected gene, determinations can be based on normalized expression levels of the gene. A gene expression level is normalized by correcting the absolute expression level of the gene (e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene as described herein) by comparing its expression to expression of a gene for which expression is not believed to be co-regulated with the gene of interest, e.g., a housekeeping gene that is constitutively expressed. Suitable genes for normalization include housekeeping genes such as the actin gene. Such normalization allows comparison of the expression level in one sample, e.g., a patient sample, with the expression level in another sample, e.g., a sample obtained from a patient known not to be afflicted with a disease or condition, or between samples obtained from different sources.

Alternatively, the expression level can be assessed as a relative expression level. To assess a relative expression level for a gene (e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene, as described herein), the level of expression of the gene is determined for 10 or more samples (preferably 50 or more samples) of different isolates of cells in which the gene is believed to be expressed, prior to assessing the level of expression of the gene in the sample of interest. The mean expression level of the gene detected in the large number of samples is determined, and this value is used as a baseline expression level for the gene. The expression level of the gene assessed in the test sample (i.e., its absolute level of expression) is divided by the mean expression value to yield a relative expression level. Such a method can identify tissues or individuals which are afflicted with a disorder associated with aberrant expression of a gene of the invention.

Preferably, the samples used in the baseline determination are generated either using cells obtained from a tissue or individual known to be afflicted with a disorder (e.g., a disorder associated with aberrant expression of one of the TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 genes) or using cells obtained from a tissue or individual known not to be afflicted with the disorder. Alternatively, levels of expression of these genes in tissues or individuals known to be or not to be afflicted with the disorder can be used to assess whether the aberrant expression of the gene is associated with the disorder (e.g., with onset of the disorder, or as a symptom of the disorder over time).

Another aspect of the invention pertains to monitoring the influence of agents (e.g., drugs or other compounds) on the expression or activity of one or more of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296 in clinical trials. These and other agents are described in further detail in the following sections.

1. Diagnostic Assays

An exemplary method for detecting the presence or absence of a polypeptide or nucleic acid of the invention in a biological sample involves obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting a polypeptide or nucleic acid (e.g., mRNA, genomic DNA) of the invention such that the presence of a polypeptide or nucleic acid of the invention is detected in the biological sample. A preferred agent for detecting mRNA or genomic DNA encoding a polypeptide of the invention is a labeled nucleic acid probe capable of hybridizing to mRNA or genomic DNA encoding a polypeptide of the invention. The nucleic acid probe can be, for example, a full-length cDNA, such as the nucleic acid of any of SEQ ID NOs: 1, 9, 17, 25, 33, 45, 53, 67, and 72, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to a mRNA or genomic DNA encoding a polypeptide of the invention. Other suitable probes for use in the diagnostic assays of the invention are described herein.

A preferred agent for detecting a polypeptide of the invention is an antibody capable of binding to a polypeptide of the invention, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab′)₂) can be used. The term “labeled”, with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently labeled streptavidin. The term “biological sample” is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method of the invention can be used to detect mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of a polypeptide of the invention include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence. In vitro techniques for detection of genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of a polypeptide of the invention include introducing into a subject a labeled antibody directed against the polypeptide. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.

In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject.

In another embodiment, the methods further involve obtaining a control biological sample from a control subject, contacting the control sample with a compound or agent capable of detecting a polypeptide of the invention or mRNA or genomic DNA encoding a polypeptide of the invention, such that the presence of the polypeptide or mRNA or genomic DNA encoding the polypeptide is detected in the biological sample, and comparing the presence of the polypeptide or mRNA or genomic DNA encoding the polypeptide in the control sample with the presence of the polypeptide or mRNA or genomic DNA encoding the polypeptide in the test sample.

The invention also encompasses kits for detecting the presence of a polypeptide or nucleic acid of the invention in a biological sample (a test sample). Such kits can be used to determine if a subject is suffering from or is at increased risk of developing a disorder associated with aberrant expression of a polypeptide of the invention (e.g., one of the disorders described in the section of this disclosure wherein the individual polypeptide of the invention is discussed). For example, the kit can comprise a labeled compound or agent capable of detecting the polypeptide or mRNA encoding the polypeptide in a biological sample and means for determining the amount of the polypeptide or mRNA in the sample (e.g., an antibody which binds the polypeptide or an oligonucleotide probe which binds to DNA or mRNA encoding the polypeptide). Kits can also include instructions for observing that the tested subject is suffering from or is at risk of developing a disorder associated with aberrant expression of the polypeptide if the amount of the polypeptide or mRNA encoding the polypeptide is above or below a normal level.

For antibody-based kits, the kit can comprise, for example: (1) a first antibody (e.g., attached to a solid support) which binds to a polypeptide of the invention; and, optionally, (2) a second, different antibody which binds to either the polypeptide or the first antibody and is conjugated to a detectable agent.

For oligonucleotide-based kits, the kit can comprise, for example: (1) an oligonucleotide, e.g., a detectably labeled oligonucleotide, which hybridizes to a nucleic acid sequence encoding a polypeptide of the invention or (2) a pair of primers useful for amplifying a nucleic acid molecule encoding a polypeptide of the invention. The kit can also comprise, e.g., a buffering agent, a preservative, or a protein stabilizing agent. The kit can also comprise components necessary for detecting the detectable agent (e.g., an enzyme or a substrate). The kit can also contain a control sample or a series of control samples which can be assayed and compared to the test sample contained. Each component of the kit is usually enclosed within an individual container and all of the various containers are within a single package along with instructions for observing whether the tested subject is suffering from or is at risk of developing a disorder associated with aberrant expression of the polypeptide.

2. Prognostic Assays

The methods described herein can furthermore be utilized as diagnostic or prognostic assays to identify subjects having or at risk of developing a disease or disorder associated with aberrant expression or activity of a polypeptide of the invention. For example, the assays described herein, such as the preceding diagnostic assays or the following assays, can be utilized to identify a subject having or at risk of developing a disorder associated with aberrant expression or activity of a polypeptide of the invention (e.g., one of the disorders described in the section of this disclosure wherein the individual polypeptide of the invention is discussed). Alternatively, the prognostic assays can be utilized to identify a subject having or at risk for developing such a disease or disorder. Thus, the present invention provides a method in which a test sample is obtained from a subject and a polypeptide or nucleic acid (e.g., mRNA, genomic DNA) of the invention is detected, wherein the presence of the polypeptide or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant expression or activity of the polypeptide. As used herein, a “test sample” refers to a biological sample obtained from a subject of interest. For example, a test sample can be a biological fluid (e.g., serum), cell sample, or tissue.

Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat a disease or disorder associated with aberrant expression or activity of a polypeptide of the invention. For example, such methods can be used to determine whether a subject can be effectively treated with a specific agent or class of agents (e.g., agents of a type which decrease activity of the polypeptide). Thus, the present invention provides methods for determining whether a subject can be effectively treated with an agent for a disorder associated with aberrant expression or activity of a polypeptide of the invention in which a test sample is obtained and the polypeptide or nucleic acid encoding the polypeptide is detected (e.g., wherein the presence of the polypeptide or nucleic acid is diagnostic for a subject that can be administered the agent to treat a disorder associated with aberrant expression or activity of the polypeptide).

The methods of the invention can also be used to detect genetic lesions or mutations in a gene of the invention, thereby determining if a subject with the lesioned gene is at risk for a disorder characterized aberrant expression or activity of a polypeptide of the invention. In preferred embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic lesion or mutation characterized by at least one of an alteration affecting the integrity of a gene encoding the polypeptide of the invention, or the mis-expression of the gene encoding the polypeptide of the invention. For example, such genetic lesions or mutations can be detected by ascertaining the existence of at least one of: 1) a deletion of one or more nucleotides from the gene; 2) an addition of one or more nucleotides to the gene; 3) a substitution of one or more nucleotides of the gene; 4) a chromosomal rearrangement of the gene; 5) an alteration in the level of a messenger RNA transcript of the gene; 6) an aberrant modification of the gene, such as of the methylation pattern of the genomic DNA; 7) the presence of a non-wild type splicing pattern of a messenger RNA transcript of the gene; 8) a non-wild type level of the protein encoded by the gene; 9) an allelic loss of the gene; and 10) an inappropriate post-translational modification of the protein encoded by the gene. As described herein, there are a large number of assay techniques known in the art which can be used for detecting lesions in a gene.

In certain embodiments, detection of the lesion involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran et al. (1988) Science 241:1077-1080; and Nakazawa et al. (1994) Proc. Natl. Acad. Sci. USA 91:360-364), the latter of which can be particularly useful for detecting point mutations in a gene (see, e.g., Abravaya et al. (1995) Nucleic Acids Res. 23:675-682). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers which specifically hybridize to the selected gene under conditions such that hybridization and amplification of the gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. PCR and/or LCR can be desirable to use as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein.

Alternative amplification methods include: self-sustained sequence replication (Guatelli et al. (1990) Proc. Natl. Acad. Sci. USA 87:1874-1878), transcriptional amplification system (Kwoh, et al. (1989) Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta Replicase (Lizardi et al. (1988) Bio/Technology 6:1197), or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers.

In an alternative embodiment, mutations in a selected gene from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, (optionally) amplified, digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicates mutations in the sample DNA. Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Pat. No. 5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.

In other embodiments, genetic mutations can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, to high density arrays containing hundreds or thousands of oligonucleotides probes (Cronin et al. (1996) Human Mutation 7:244-255; Kozal et al. (1996) Nature Medicine 2:753-759). For example, genetic mutations can be identified in two-dimensional arrays containing light-generated DNA probes as described in Cronin et al., supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This step is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.

In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the selected gene and detect mutations by comparing the sequence of the sample nucleic acids with the corresponding wild-type (control) sequence. Examples of sequencing reactions include those based on techniques developed by Maxim and Gilbert ((1977) Proc. Natl. Acad. Sci. USA 74:560) or Sanger ((1977) Proc. Natl. Acad. Sci. USA 74:5463). It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays ((1995) Bio/Techniques 19:448), including sequencing by mass spectrometry (see, e.g., PCT Publication No. WO 94/16101; Cohen et al. (1996) Adv. Chromatogr. 36:127-162; and Griffin et al. (1993) Appl. Biochem. Biotechnol. 38:147-159).

Other methods for detecting mutations in a selected gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes (Myers et al. (1985) Science 230:1242). In general, the technique of mismatch cleavage entails providing heteroduplexes formed by hybridizing (labeled) RNA or DNA containing the wild-type sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent which cleaves single-stranded regions of the duplex such as which will exist due to base pair mismatches between the control and sample strands. RNA/DNA duplexes can be treated with RNase to digest mismatched regions, and DNA/DNA hybrids can be treated with S1 nuclease to digest mismatched regions.

In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion of the mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. See, e.g., Cotton et al. (1988) Proc. Natl. Acad. Sci. USA 85:4397; Saleeba et al. (1992) Methods Enzymol 217:286-295. In a preferred embodiment, the control DNA or RNA can be labeled for detection.

In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called DNA mismatch repair enzymes) in defined systems for detecting and mapping point mutations in cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches (Hsu et al. (1994) Carcinogenesis 15:1657-1662). According to an exemplary embodiment, a probe based on a selected sequence, e.g., a wild-type sequence, is hybridized to a cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage, products, if any, can be detected from electrophoresis protocols or the like. See, e.g., U.S. Pat. No. 5,459,039.

In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in genes. For example, single strand conformation polymorphism (SSCP) can be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids (Orita et al. (1989) Proc. Natl. Acad. Sci. USA 86:2766; see also Cotton (1993) Mutat. Res. 285:125-144; Hayashi (1992) Genet. Anal. Tech. Appl. 9:73-79). Single-stranded DNA fragments of sample and control nucleic acids will be denatured and allowed to re-nature. The secondary structure of single-stranded nucleic acids varies according to sequence, and the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments can be labeled or detected with labeled probes. The sensitivity of the assay can be enhanced by using RNA (rather than DNA), in which the secondary structure is more sensitive to a change in sequence. In a preferred embodiment, the subject method utilizes heteroduplexes analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility (Keen et al. (1991) Trends Genet. 7:5).

In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE) (Myers et al. (1985) Nature 313:495). When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a ‘GC clamp’ of approximately 40 base pairs of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA (Rosenbaum and Reissner (1987) Biophys. Chem. 265:12753).

Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, oligonucleotide primers can be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions which permit hybridization only if a perfect match is found (Saiki et al. (1986) Nature 324:163); Saiki et al. (1989) Proc. Natl. Acad. Sci. USA 86:6230). Such allele specific oligonucleotides are hybridized to PCR amplified target DNA or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA.

Alternatively, allele specific amplification technology which depends on selective PCR amplification can be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification can carry the mutation of interest in the center of the molecule (so that amplification depends on differential hybridization; Gibbs et al. (1989) Nucleic Acids Res. 17:2437-2448) or at the extreme 3′ end of one primer where, under appropriate conditions, mismatching can prevent or reduce polymerase extension (Prossner (1993) Tibtech 11:238). In addition, it can be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection (Gasparini et al. (1992) Mol. Cell Probes 6:1). Amplification can also be performed using Taq ligase for amplification (Barany (1991) Proc. Natl. Acad. Sci. USA 88:189). In such cases, ligation will occur only if there is a perfect match at the 3′ end of the 5′ sequence making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.

The methods described herein can be performed, for example, using pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which can be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving a gene encoding a polypeptide of the invention. Furthermore, any cell type or tissue, preferably peripheral blood leukocytes, in which the polypeptide of the invention is expressed can be utilized in the prognostic assays described herein.

3. Pharmacogenomics

Agents, or modulators which have a stimulatory or inhibitory effect on activity or expression of a polypeptide of the invention as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) disorders associated with aberrant activity of the polypeptide. In conjunction with such treatment, the pharmacogenomics (i.e., the study of the relationship between an individual's genotype and that individual's response to a foreign compound or drug) of the individual may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, the pharmacogenomics of the individual permits the selection of effective agents (e.g., drugs) for prophylactic or therapeutic treatments based on a consideration of the individual's genotype. Such pharmacogenomics can further be used to determine appropriate dosages and therapeutic regimens. Accordingly, the activity of a polypeptide of the invention, expression of a nucleic acid of the invention, or mutation content of a gene of the invention in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual.

Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See, e.g., Linder (1997) Clin. Chem. 43(2):254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body are referred to as “altered drug action.” Genetic conditions transmitted as single factors altering the way the body acts on drugs are referred to as “altered drug metabolism”. These pharmacogenetic conditions can occur either as rare defects or as polymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

As an illustrative embodiment, the activity of drug metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymorphisms of drug metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYP2C19) has provided an explanation as to why some patients do not obtain the expected drug effects or show exaggerated drug response and serious toxicity after taking the standard and safe dose of a drug. These polymorphisms are expressed in two phenotypes in the population, the extensive metabolizer (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. For example, the gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quite frequently experience exaggerated drug response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, a PM will show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. The other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.

Thus, the activity of a polypeptide of the invention, expression of a nucleic acid encoding the polypeptide, or mutation content of a gene encoding the polypeptide in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual. In addition, pharmacogenetic studies can be used to apply genotyping of polymorphic alleles encoding drug-metabolizing enzymes to the identification of an individual's drug responsiveness phenotype. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with a modulator of activity or expression of the polypeptide, such as a modulator identified by one of the exemplary screening assays described herein.

4. Monitoring of Effects During Clinical Trials

Monitoring the influence of agents (e.g., drug compounds) on the expression or activity of a polypeptide of the invention (e.g., the ability to modulate aberrant cell proliferation chemotaxis, and/or differentiation) can be applied not only in basic drug screening, but also in clinical trials. For example, the effectiveness of an agent, as determined by a screening assay as described herein, to increase gene expression, protein levels, or protein activity, can be monitored in clinical trials of subjects exhibiting decreased gene expression, protein levels, or protein activity. Alternatively, the effectiveness of an agent, as determined by a screening assay, to decrease gene expression, protein levels or protein activity, can be monitored in clinical trials of subjects exhibiting increased gene expression, protein levels, or protein activity. In such clinical trials, expression or activity of a polypeptide of the invention and preferably, that of other polypeptide that have been implicated in for example, a cellular proliferation disorder, can be used as a marker of the immune responsiveness of a particular cell.

For example, and not by way of limitation, genes, including those of the invention, that are modulated in cells by treatment with an agent (e.g., compound, drug or small molecule) which modulates activity or expression of a polypeptide of the invention (e.g., as identified in a screening assay described herein) can be identified. Thus, to study the effect of agents on cellular proliferation disorders, for example, in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of a gene of the invention and other genes implicated in the disorder. The levels of gene expression (i.e., a gene expression pattern) can be quantified by Northern blot analysis or RT-PCR, as described herein, or alternatively by measuring the amount of protein produced, by one of the methods as described herein, or by measuring the levels of activity of a gene of the invention or other genes. In this way, the gene expression pattern can serve as a marker, indicative of the physiological response of the cells to the agent. Accordingly, this response state can be determined before, and at various points during, treatment of the individual with the agent.

In a preferred embodiment, the present invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate identified by the screening assays described herein) comprising the steps of (i) obtaining a pre-administration sample from a subject prior to administration of the agent; (ii) detecting the level of the polypeptide or nucleic acid of the invention in the pre-administration sample; (iii) obtaining one or more post-administration samples from the subject; (iv) detecting the level the of the polypeptide or nucleic acid of the invention in the post-administration samples; (v) comparing the level of the polypeptide or nucleic acid of the invention in the pre-administration sample with the level of the polypeptide or nucleic acid of the invention in the post-administration sample or samples; and (vi) altering the administration of the agent to the subject accordingly. For example, increased administration of the agent can be desirable to increase the expression or activity of the polypeptide to higher levels than detected, i.e., to increase the effectiveness of the agent. Alternatively, decreased administration of the agent can be desirable to decrease expression or activity of the polypeptide to lower levels than detected, i.e., to decrease the effectiveness of the agent.

C. Methods of Treatment

The present invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant expression or activity of a polypeptide of the invention and/or in which the polypeptide of the invention is involved. Disorders characterized by aberrant expression or activity of the polypeptides of the invention are described elsewhere in this disclosure.

1. Prophylactic Methods

In one aspect, the invention provides a method for preventing in a subject, a disease or condition associated with an aberrant expression or activity of a polypeptide of the invention, by administering to the subject an agent which modulates expression or at least one activity of the polypeptide. Subjects at risk for a disease which is caused or contributed to by aberrant expression or activity of a polypeptide of the invention can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the aberrance, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending on the type of aberrance, for example, an agonist or antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein.

2. Therapeutic Methods

Another aspect of the invention pertains to methods of modulating expression or activity of a polypeptide of the invention for therapeutic purposes. The modulatory method of the invention involves contacting a cell with an agent that modulates one or more of the activities of the polypeptide. An agent that modulates activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring cognate ligand of the polypeptide, a peptide, a peptidomimetic, or other small molecule. In one embodiment, the agent stimulates one or more of the biological activities of the polypeptide. Examples of such stimulatory agents include the active polypeptide of the invention and a nucleic acid molecule encoding the polypeptide of the invention that has been introduced into the cell. In another embodiment, the agent inhibits one or more of the biological activities of the polypeptide of the invention. Examples of such inhibitory agents include antisense nucleic acid molecules and antibodies. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the present invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant expression or activity of a polypeptide of the invention. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up-regulates or down-regulates) expression or activity. In another embodiment, the method involves administering a polypeptide of the invention or a nucleic acid molecule of the invention as therapy to compensate for reduced or aberrant expression or activity of the polypeptide.

Stimulation of activity is desirable in situations in which activity or expression is abnormally low or down-regulated and/or in which increased activity is likely to have a beneficial effect. Conversely, inhibition of activity is desirable in situations in which activity or expression is abnormally high or up-regulated and/or in which decreased activity is likely to have a beneficial effect.

The contents of all references, patents, and published patent applications cited throughout this application are hereby incorporated by reference.

Deposit of Clones

Each of these deposits was made merely as a convenience to those of skill in the art. These deposits are not an admission that a deposit is required under 35 U.S.C. §112.

Clone EpT202, encoding human TANGO 202 was deposited with the American Type Culture Collection (ATCC®, 10801 University Boulevard, Manassas, Va. 20110-2209) on Apr. 21, 1999 and was assigned Accession Number 207219. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpTm202, encoding murine TANGO 202 was deposited with ATCC® on Apr. 21, 1999 and was assigned (composite) Accession Number 207221. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT234, encoding human TANGO 234 was deposited with ATTC® on Apr. 2, 1999 and was assigned Accession Number 207184. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT265, encoding human TANGO 265 was deposited with ATCC® on Apr. 28, 1999 and was assigned Accession Number 207228. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT273, encoding human TANGO 273 was deposited with ATCC® on Apr. 2, 1999 and was assigned Accession Number 207185. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpTm273, encoding murine TANGO 273 was deposited with ATCC® on Apr. 2, 1999 and was assigned (composite) Accession Number 207221. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT286, encoding human TANGO 286 was deposited with ATTC® on Apr. 20, 1999 and was assigned (composite) Accession Number 207220. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT294, encoding human TANGO 294 was deposited with ATCC® on Apr. 20, 1999 and was assigned (composite) Accession Number 207220. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clone EpT296, encoding human INTERCEPT 296 was deposited with ATTC® on Apr. 20, 1999 and was assigned (composite) Accession Number 207220. This deposit will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

Clones containing cDNA molecules encoding human TANGO 286, human TANGO 294, and INTERCEPT 296 were deposited with ATTC® on Apr. 21, 1999 as Accession Number 207220, as part of a composite deposit representing a mixture of five strains, each carrying one recombinant plasmid harboring a particular cDNA clone.

To distinguish the strains and isolate a strain harboring a particular cDNA clone, an aliquot of the mixture is streaked out to single colonies on nutrient medium (e.g., LB plates) supplemented with 100 mg/ml ampicillin, single colonies are grown, and then plasmid DNA is extracted using a standard mini-preparation procedure. Next, a sample of the DNA mini-preparation is digested with a combination of the restriction enzymes SalI, NotI, and DraII and the resulting products are resolved on a 0.8% agarose gel using standard DNA electrophoresis conditions. This digestion procedure liberates fragments as follows:

1. human TANGO 286 (clone EpT286): 1.85 kB and 0.1 kB (human TANGO 286 has a DraII cut site at about base pair 1856).

2. human TANGO 294 (clone EpT294): 1.4 kB and 0.6 kB (human TANGO 294 has a DraII cut site at about base pair 1447).

3. human INTERCEPT 296 (clone EpT296): 0.4 kB, 1.6 kB, and 0.1 kB (human INTERCEPT 296 has DraII cut sites at about base pair 410 and at about base pair 1933).

The identity of the strains can be inferred from the fragments liberated.

Clones containing cDNA molecules encoding mouse TANGO 202 and mouse TANGO 273 were deposited with ATCC® on Apr. 21, 1999 and were assigned Accession Number 207221, as part of a composite deposit representing a mixture of five strains, each carrying one recombinant plasmid harboring a particular cDNA clone.

To distinguish the strains and isolate a strain harboring a particular cDNA clone, an aliquot of the mixture is streaked out to single colonies on nutrient medium (e.g., LB plates) supplemented with 100 mg/ml ampicillin, single colonies are grown, and then plasmid DNA is extracted using a standard mini-preparation procedure. Next, a sample of the DNA mini-preparation is digested with a combination of the restriction enzymes Sal I, Not I, and Apa I, and the resultant products are resolved on a 0.8% agarose gel using standard DNA electrophoresis conditions. This digestion procedure liberates fragments as follows:

1. mouse TANGO 202 (clone EpTm202): 3.5 kB and 1.4 kB (mouse TANGO 202 has a Apa I cut site at about base pair 3519).

2. mouse TANGO 273 (clone EpTm273): 0.3 kB and 2.6 kB (mouse TANGO 273 has a Apa I cut site at about base pair 298).

The identity of the strains can be inferred from the fragments liberated.

Human TANGO 202, human TANGO 234, human TANGO 265, and human TANGO 273 were each deposited as single deposits. Their clone names, deposit dates, and accession numbers are as follows:

1. human TANGO 202: clone EpT202 was deposited with ATCC® on Apr. 21, 1999, and was assigned Accession Number 207219.

2. human TANGO 234: clone EpT234 was deposited with ATCC® on Apr. 2, 1999, and was assigned Accession Number 207184.

3. human TANGO 265: clone EpT265 was deposited with ATCC® on Apr. 28, 1999, and was assigned Accession Number 207228.

4. human TANGO 273: clone EpT273 was deposited with ATCC® on Apr. 2, 1999, and was assigned Accession Number 207185.

All publications, patents, and patent applications referenced in this specification are incorporated by reference into the specification to the same extent as if each individual publication, patent, or patent application had been specifically and individually indicated to be incorporated herein by reference.

Equivalents

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

79 1 1656 DNA Homo sapiens 1 gtcgacccac gcgtccgccc acgcgtccgg cccatggcgc cgcccgccgc ccgcctcgcc 60 ctgctctccg ccgcggcgct cacgctggcg gcccggcccg cgcctagccc cggcctcggc 120 cccggacccg agtgtttcac agccaatggt gcggattata ggggaacaca gaactggaca 180 gcactacaag gcgggaagcc atgtctgttt tggaacgaga ctttccagca tccatacaac 240 actctgaaat accccaacgg ggaggggggc ctgggtgagc acaactattg cagaaatcca 300 gatggagacg tgagcccctg gtgctatgtg gcagagcacg aggatggtgt ctactggaag 360 tactgtgaga tacctgcttg ccagatgcct ggaaaccttg gctgctacaa ggatcatgga 420 aacccacctc ctctaactgg caccagtaaa acgtccaaca aactcaccat acaaacttgc 480 atcagttttt gtcggagtca gaggttcaag tttgctggga tggagtcagg ctatgcttgc 540 ttctgtggaa acaatcctga ttactggaag tacggggagg cagccagtac cgaatgcaac 600 agcgtctgct tcggggatca cacccaaccc tgtggtggcg atggcaggat catcctcttt 660 gatactctcg tgggcgcctg cggtgggaac tactcagcca tgtcttctgt ggtctattcc 720 cctgacttcc ccgacaccta tgccacgggg agggtctgct actggaccat ccgggttccg 780 ggggcctccc acatccactt cagcttcccc ctatttgaca tcagggactc ggcggacatg 840 gtggagcttc tggatggcta cacccaccgt gtcctagccc gcttccacgg gaggagccgc 900 ccacctctgt ccttcaacgt ctctctggac ttcgtcatct tgtatttctt ctctgatcgc 960 atcaatcagg cccagggatt tgctgtttta taccaagccg tcaaggaaga actgccacag 1020 gagaggcccg ctgtcaacca gacggtggcc gaggtgatca cggagcaggc caacctcagt 1080 gtcagcgctg cccggtcctc caaagtcctc tatgtcatca ccaccagccc cagccaccca 1140 cctcagactg tcccaggtag caattcctgg gcgccaccca tgggggctgg aagccacaga 1200 gttgaaggat ggacagtcta tggtctggca actctcctca tcctcacagt cacagccatt 1260 gtagcaaaga tacttctgca cgtcacattc aaatcccatc gtgttcctgc ttcaggggac 1320 cttagggatt gtcatcaacc agggacttcg ggggaaatct ggagcatttt ttacaagcct 1380 tccacttcaa tttccatctt taagaagaaa ctcaagggtc agagtcaaca agatgaccgc 1440 aatccccttg tgagtgacta aaaaccccac tgtgcctagg acttgaggtc cctctttgag 1500 ctcaaggctg ccgtggtcaa cctctcctgt ggttcttctc tgacagactc ttccctcctc 1560 tccctctgcc tcggcctctt cggggaaacc ctcctcctac agactaggaa gaggcacctg 1620 ctgccagggc aggcagagcc tggattcctc ctgctt 1656 2 1425 DNA Homo sapiens 2 atggcgccgc ccgccgcccg cctcgccctg ctctccgccg cggcgctcac gctggcggcc 60 cggcccgcgc ctagccccgg cctcggcccc ggacccgagt gtttcacagc caatggtgcg 120 gattataggg gaacacagaa ctggacagca ctacaaggcg ggaagccatg tctgttttgg 180 aacgagactt tccagcatcc atacaacact ctgaaatacc ccaacgggga ggggggcctg 240 ggtgagcaca actattgcag aaatccagat ggagacgtga gcccctggtg ctatgtggca 300 gagcacgagg atggtgtcta ctggaagtac tgtgagatac ctgcttgcca gatgcctgga 360 aaccttggct gctacaagga tcatggaaac ccacctcctc taactggcac cagtaaaacg 420 tccaacaaac tcaccataca aacttgcatc agtttttgtc ggagtcagag gttcaagttt 480 gctgggatgg agtcaggcta tgcttgcttc tgtggaaaca atcctgatta ctggaagtac 540 ggggaggcag ccagtaccga atgcaacagc gtctgcttcg gggatcacac ccaaccctgt 600 ggtggcgatg gcaggatcat cctctttgat actctcgtgg gcgcctgcgg tgggaactac 660 tcagccatgt cttctgtggt ctattcccct gacttccccg acacctatgc cacggggagg 720 gtctgctact ggaccatccg ggttccgggg gcctcccaca tccacttcag cttcccccta 780 tttgacatca gggactcggc ggacatggtg gagcttctgg atggctacac ccaccgtgtc 840 ctagcccgct tccacgggag gagccgccca cctctgtcct tcaacgtctc tctggacttc 900 gtcatcttgt atttcttctc tgatcgcatc aatcaggccc agggatttgc tgttttatac 960 caagccgtca aggaagaact gccacaggag aggcccgctg tcaaccagac ggtggccgag 1020 gtgatcacgg agcaggccaa cctcagtgtc agcgctgccc ggtcctccaa agtcctctat 1080 gtcatcacca ccagccccag ccacccacct cagactgtcc caggtagcaa ttcctgggcg 1140 ccacccatgg gggctggaag ccacagagtt gaaggatgga cagtctatgg tctggcaact 1200 ctcctcatcc tcacagtcac agccattgta gcaaagatac ttctgcacgt cacattcaaa 1260 tcccatcgtg ttcctgcttc aggggacctt agggattgtc atcaaccagg gacttcgggg 1320 gaaatctgga gcatttttta caagccttcc acttcaattt ccatctttaa gaagaaactc 1380 aagggtcaga gtcaacaaga tgaccgcaat ccccttgtga gtgac 1425 3 475 PRT Homo sapiens 3 Met Ala Pro Pro Ala Ala Arg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala Ala Arg Pro Ala Pro Ser Pro Gly Leu Gly Pro Gly Pro 20 25 30 Glu Cys Phe Thr Ala Asn Gly Ala Asp Tyr Arg Gly Thr Gln Asn Trp 35 40 45 Thr Ala Leu Gln Gly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe 50 55 60 Gln His Pro Tyr Asn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu 65 70 75 80 Gly Glu His Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp 85 90 95 Cys Tyr Val Ala Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu 100 105 110 Ile Pro Ala Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His 115 120 125 Gly Asn Pro Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu 130 135 140 Thr Ile Gln Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe 145 150 155 160 Ala Gly Met Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp 165 170 175 Tyr Trp Lys Tyr Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys 180 185 190 Phe Gly Asp His Thr Gln Pro Cys Gly Gly Asp Gly Arg Ile Ile Leu 195 200 205 Phe Asp Thr Leu Val Gly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ser 210 215 220 Ser Val Val Tyr Ser Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg 225 230 235 240 Val Cys Tyr Trp Thr Ile Arg Val Pro Gly Ala Ser His Ile His Phe 245 250 255 Ser Phe Pro Leu Phe Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu 260 265 270 Leu Asp Gly Tyr Thr His Arg Val Leu Ala Arg Phe His Gly Arg Ser 275 280 285 Arg Pro Pro Leu Ser Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr 290 295 300 Phe Phe Ser Asp Arg Ile Asn Gln Ala Gln Gly Phe Ala Val Leu Tyr 305 310 315 320 Gln Ala Val Lys Glu Glu Leu Pro Gln Glu Arg Pro Ala Val Asn Gln 325 330 335 Thr Val Ala Glu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala 340 345 350 Ala Arg Ser Ser Lys Val Leu Tyr Val Ile Thr Thr Ser Pro Ser His 355 360 365 Pro Pro Gln Thr Val Pro Gly Ser Asn Ser Trp Ala Pro Pro Met Gly 370 375 380 Ala Gly Ser His Arg Val Glu Gly Trp Thr Val Tyr Gly Leu Ala Thr 385 390 395 400 Leu Leu Ile Leu Thr Val Thr Ala Ile Val Ala Lys Ile Leu Leu His 405 410 415 Val Thr Phe Lys Ser His Arg Val Pro Ala Ser Gly Asp Leu Arg Asp 420 425 430 Cys His Gln Pro Gly Thr Ser Gly Glu Ile Trp Ser Ile Phe Tyr Lys 435 440 445 Pro Ser Thr Ser Ile Ser Ile Phe Lys Lys Lys Leu Lys Gly Gln Ser 450 455 460 Gln Gln Asp Asp Arg Asn Pro Leu Val Ser Asp 465 470 475 4 19 PRT Homo sapiens 4 Met Ala Pro Pro Ala Ala Arg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala 5 456 PRT Homo sapiens 5 Ala Arg Pro Ala Pro Ser Pro Gly Leu Gly Pro Gly Pro Glu Cys Phe 1 5 10 15 Thr Ala Asn Gly Ala Asp Tyr Arg Gly Thr Gln Asn Trp Thr Ala Leu 20 25 30 Gln Gly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His Pro 35 40 45 Tyr Asn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu His 50 55 60 Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr Val 65 70 75 80 Ala Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro Ala 85 90 95 Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn Pro 100 105 110 Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile Gln 115 120 125 Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly Met 130 135 140 Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp Tyr Trp Lys 145 150 155 160 Tyr Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys Phe Gly Asp 165 170 175 His Thr Gln Pro Cys Gly Gly Asp Gly Arg Ile Ile Leu Phe Asp Thr 180 185 190 Leu Val Gly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ser Ser Val Val 195 200 205 Tyr Ser Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg Val Cys Tyr 210 215 220 Trp Thr Ile Arg Val Pro Gly Ala Ser His Ile His Phe Ser Phe Pro 225 230 235 240 Leu Phe Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu Leu Asp Gly 245 250 255 Tyr Thr His Arg Val Leu Ala Arg Phe His Gly Arg Ser Arg Pro Pro 260 265 270 Leu Ser Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe Ser 275 280 285 Asp Arg Ile Asn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala Val 290 295 300 Lys Glu Glu Leu Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Val Ala 305 310 315 320 Glu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala Arg Ser 325 330 335 Ser Lys Val Leu Tyr Val Ile Thr Thr Ser Pro Ser His Pro Pro Gln 340 345 350 Thr Val Pro Gly Ser Asn Ser Trp Ala Pro Pro Met Gly Ala Gly Ser 355 360 365 His Arg Val Glu Gly Trp Thr Val Tyr Gly Leu Ala Thr Leu Leu Ile 370 375 380 Leu Thr Val Thr Ala Ile Val Ala Lys Ile Leu Leu His Val Thr Phe 385 390 395 400 Lys Ser His Arg Val Pro Ala Ser Gly Asp Leu Arg Asp Cys His Gln 405 410 415 Pro Gly Thr Ser Gly Glu Ile Trp Ser Ile Phe Tyr Lys Pro Ser Thr 420 425 430 Ser Ile Ser Ile Phe Lys Lys Lys Leu Lys Gly Gln Ser Gln Gln Asp 435 440 445 Asp Arg Asn Pro Leu Val Ser Asp 450 455 6 373 PRT Homo sapiens 6 Ala Arg Pro Ala Pro Ser Pro Gly Leu Gly Pro Gly Pro Glu Cys Phe 1 5 10 15 Thr Ala Asn Gly Ala Asp Tyr Arg Gly Thr Gln Asn Trp Thr Ala Leu 20 25 30 Gln Gly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His Pro 35 40 45 Tyr Asn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu His 50 55 60 Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr Val 65 70 75 80 Ala Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro Ala 85 90 95 Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn Pro 100 105 110 Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile Gln 115 120 125 Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly Met 130 135 140 Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp Tyr Trp Lys 145 150 155 160 Tyr Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys Phe Gly Asp 165 170 175 His Thr Gln Pro Cys Gly Gly Asp Gly Arg Ile Ile Leu Phe Asp Thr 180 185 190 Leu Val Gly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ser Ser Val Val 195 200 205 Tyr Ser Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg Val Cys Tyr 210 215 220 Trp Thr Ile Arg Val Pro Gly Ala Ser His Ile His Phe Ser Phe Pro 225 230 235 240 Leu Phe Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu Leu Asp Gly 245 250 255 Tyr Thr His Arg Val Leu Ala Arg Phe His Gly Arg Ser Arg Pro Pro 260 265 270 Leu Ser Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe Ser 275 280 285 Asp Arg Ile Asn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala Val 290 295 300 Lys Glu Glu Leu Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Val Ala 305 310 315 320 Glu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala Arg Ser 325 330 335 Ser Lys Val Leu Tyr Val Ile Thr Thr Ser Pro Ser His Pro Pro Gln 340 345 350 Thr Val Pro Gly Ser Asn Ser Trp Ala Pro Pro Met Gly Ala Gly Ser 355 360 365 His Arg Val Glu Gly 370 7 23 PRT Homo sapiens 7 Trp Thr Val Tyr Gly Leu Ala Thr Leu Leu Ile Leu Thr Val Thr Ala 1 5 10 15 Ile Val Ala Lys Ile Leu Leu 20 8 60 PRT Homo sapiens 8 His Val Thr Phe Lys Ser His Arg Val Pro Ala Ser Gly Asp Leu Arg 1 5 10 15 Asp Cys His Gln Pro Gly Thr Ser Gly Glu Ile Trp Ser Ile Phe Tyr 20 25 30 Lys Pro Ser Thr Ser Ile Ser Ile Phe Lys Lys Lys Leu Lys Gly Gln 35 40 45 Ser Gln Gln Asp Asp Arg Asn Pro Leu Val Ser Asp 50 55 60 9 4628 DNA Homo sapiens 9 gcggccgctc gcgatctaga actagtaatg atgctgcctc aaaactcgtg gcatattgat 60 tttggaagat gctgctgtca tcagaacctt ttctctgctg tggtaacttg catcctgctc 120 ctgaattcct gctttctcat cagcagtttt aatggaacag atttggagtt gaggctggtc 180 aatggagacg gtccctgctc tgggacagtg gaggtgaaat tccagggaca gtgggggact 240 gtgtgtgatg atgggtggaa cactactgcc tcaactgtcg tgtgcaaaca gcttggatgt 300 ccattttctt tcgccatgtt tcgttttgga caagccgtga ctagacatgg aaaaatttgg 360 cttgatgatg tttcctgtta tggaaatgag tcagctctct gggaatgtca acaccgggaa 420 tggggaagcc ataactgtta tcatggagaa gatgttggtg tgaactgtta tggtgaagcc 480 aatctgggtt tgaggctagt ggatggaaac aactcctgtt cagggagagt ggaggtgaaa 540 ttccaagaaa ggtgggggac tatatgtgat gatgggtgga acttgaatac tgctgccgtg 600 gtgtgcaggc aactaggatg tccatcttct tttatttctt ctggagttgt taatagccct 660 gctgtattgc gccccatttg gctggatgac attttatgcc aggggaatga gttggcactc 720 tggaattgca gacatcgtgg atggggaaat catgactgca gtcacaatga ggatgtcaca 780 ttaacttgtt atgatagtag tgatcttgaa ctaaggcttg taggtggaac taaccgctgt 840 atggggagag tagagctgaa aatccaagga aggtggggga ccgtatgcca ccataagtgg 900 aacaatgctg cagctgatgt cgtatgcaag cagttgggat gtggaaccgc acttcacttc 960 gctggcttgc ctcatttgca gtcagggtct gatgttgtat ggcttgatgg tgtctcctgc 1020 tccggtaatg aatcttttct ttgggactgc agacattccg gaaccgtcaa ttttgactgt 1080 cttcatcaaa acgatgtgtc tgtgatctgc tcagatggag cagatttgga actgcgacta 1140 gcagatggaa gtaacaattg ttcagggaga gtagaggtga gaattcatga acagtggtgg 1200 acaatatgtg accagaactg gaagaatgaa caagcccttg tggtttgtaa gcagctagga 1260 tgtccgttca gcgtctttgg cagtcgtcgt gctaaaccta gtaatgaagc tagagacatt 1320 tggataaaca gcatatcttg cactgggaat gagtcagctc tctgggactg cacatatgat 1380 ggaaaagcaa agcgaacatg cttccgaaga tcagatgctg gagtaatttg ttctgataag 1440 gcagatctgg acctaaggct tgtcggggct catagcccct gttatgggag attggaggtg 1500 aaataccaag gagagtgggg gactgtgtgt catgacagat ggagcacaag gaatgcagct 1560 gttgtgtgta aacaattggg atgtggaaag cctatgcatg tgtttggtat gacctatttt 1620 aaagaagcat caggacctat ttggctggat gacgtttctt gcattggaaa tgagtcaaat 1680 atctgggact gtgaacacag tggatgggga aagcataatt gtgtacacag agaggatgtg 1740 attgtaacct gctcaggtga tgcaacatgg ggcctgaggc tggtgggcgg cagcaaccgc 1800 tgctcgggaa gactggaggt gtactttcaa ggacggtggg gcacagtgtg tgatgacggc 1860 tggaacagta aagctgcagc tgtggtgtgt agccagctgg actgcccatc ttctatcatt 1920 ggcatgggtc tgggaaacgc ttctacagga tatggaaaaa tttggctcga tgatgtttcc 1980 tgtgatggag atgagtcaga tctctggtca tgcaggaaca gtgggtgggg aaataatgac 2040 tgcagtcaca gtgaagatgt tggagtgatc tgttctgatg catcggatat ggagctgagg 2100 cttgtgggtg gaagcagcag gtgtgctgga aaagttgagg tgaatgtcca gggtgccgtg 2160 ggaattctgt gtgctaatgg ctggggaatg aacattgctg aagttgtttg caggcaactt 2220 gaatgtgggt ctgcaatcag ggtctccaga gagcctcatt tcacagaaag aacattacac 2280 atcttaatgt cgaattctgg ctgcactgga ggggaagcct ctctctggga ttgtatacga 2340 tgggagtgga aacagactgc gtgtcattta aatatggaag caagtttgat ctgctcagcc 2400 cacaggcagc ccaggctggt tggagctgat atgccctgct ctggacgtgt tgaagtgaaa 2460 catgcagaca catggcgctc tgtctgtgat tctgatttct ctcttcatgc tgccaatgtg 2520 ctgtgcagag aattaaattg tggagatgcc atatctcttt ctgtgggaga tcactttgga 2580 aaagggaatg gtctaacttg ggccgaaaag ttccagtgtg aagggagtga aactcacctt 2640 gcattatgcc ccattgttca acatccggaa gacacttgta tccacagcag agaagttgga 2700 gttgtctgtt cccgatatac agatgtccga cttgtgaatg gcaaatccca gtgtgacggg 2760 caagtggaga tcaacgtgct tggacactgg ggctcactgt gtgacaccca ctgggaccca 2820 gaagatgccc gtgttctatg cagacagctc agctgtggga ctgctctctc aaccacagga 2880 ggaaaatata ttggagaaag aagtgttcgt gtgtggggac acaggtttca ttgcttaggg 2940 aatgagtcac ttctggataa ctgtcaaatg acagttcttg gagcacctcc ctgtatccat 3000 ggaaatactg tctctgtgat ctgcacagga agcctgaccc agccactgtt tccatgcctc 3060 gcaaatgtat ctgacccata tttgtctgca gttccagagg gcagtgcttt gatctgctta 3120 gaggacaaac ggctccgcct agtggatggg gacagccgct gtgccgggag agtagagatc 3180 tatcacgacg gcttctgggg caccatctgt gatgacggct gggacctgag cgatgcccac 3240 gtggtgtgtc aaaagctggg ctgtggagtg gccttcaatg ccacggtctc tgctcacttt 3300 ggggaggggt cagggcccat ctggctggat gacctgaact gcacaggaac ggagtcccac 3360 ttgtggcagt gcccttcccg cggctggggg cagcacgact gcaggcacaa ggaggacgca 3420 ggggtcatct gctcagaatt cacagccttg aggctctaca gtgaaactga aacagagagc 3480 tgtgctggga gattggaagt cttctataac gggacctggg gcagcgtcgg caggaggaac 3540 atcaccacag ccatagcagg cattgtgtgc aggcagctgg gctgtgggga gaatggagtt 3600 gtcagcctcg cccctttatc taagacaggc tctggtttca tgtgggtgga tgacattcag 3660 tgtcctaaaa cgcatatctc catatggcag tgcctgtctg ccccatggga gcgaagaatc 3720 tccagcccag cagaagagac ctggatcaca tgtgaagata gaataagagt gcgtggagga 3780 gacaccgagt gctctgggag agtggagatc tggcacgcag gctcctgggg cacagtgtgt 3840 gatgactcct gggacctggc cgaggcggaa gtggtgtgtc agcagctggg ctgtggctct 3900 gctctggctg ccctgaggga cgcttcgttt ggccagggaa ctggaaccat ctggttggat 3960 gacatgcggt gcaaaggaaa tgagtcattt ctatgggact gtcacgccaa accctgggga 4020 cagagtgact gtggacacaa ggaagatgct ggcgtgaggt gctctggaca gtcgctgaaa 4080 tcactgaatg cctcctcagg tcatttagca cttattttat ccagtatctt tgggctcctt 4140 ctcctggttc tgtttattct atttctcacg tggtgccgag ttcagaaaca aaaacatctg 4200 cccctcagag tttcaaccag aaggaggggt tctctcgagg agaatttatt ccatgagatg 4260 gagacctgcc tcaagagaga ggacccacat gggacaagaa cctcagatga cacccccaac 4320 catggttgtg aagatgctag cgacacatcg ctgttgggag ttcttcctgc ctctgaagcc 4380 acaaaatgac tttagacttc cagggctcac cagatcaacc tctaaatatc tttgaaggag 4440 acaacaactt ttaaatgaat aaagaggaag tcaagttgcc ctatggaaaa cttgtccaaa 4500 taacatttct tgaacaatag gagaacagct aaattgataa agactggtga taataaaaat 4560 tgaattatgt atatcactgt taaaaaaaaa aaaaaaaaaa aaaaaaaaaa acggacgcgt 4620 gggtcgac 4628 10 4359 DNA Homo sapiens 10 atgatgctgc ctcaaaactc gtggcatatt gattttggaa gatgctgctg tcatcagaac 60 cttttctctg ctgtggtaac ttgcatcctg ctcctgaatt cctgctttct catcagcagt 120 tttaatggaa cagatttgga gttgaggctg gtcaatggag acggtccctg ctctgggaca 180 gtggaggtga aattccaggg acagtggggg actgtgtgtg atgatgggtg gaacactact 240 gcctcaactg tcgtgtgcaa acagcttgga tgtccatttt ctttcgccat gtttcgtttt 300 ggacaagccg tgactagaca tggaaaaatt tggcttgatg atgtttcctg ttatggaaat 360 gagtcagctc tctgggaatg tcaacaccgg gaatggggaa gccataactg ttatcatgga 420 gaagatgttg gtgtgaactg ttatggtgaa gccaatctgg gtttgaggct agtggatgga 480 aacaactcct gttcagggag agtggaggtg aaattccaag aaaggtgggg gactatatgt 540 gatgatgggt ggaacttgaa tactgctgcc gtggtgtgca ggcaactagg atgtccatct 600 tcttttattt cttctggagt tgttaatagc cctgctgtat tgcgccccat ttggctggat 660 gacattttat gccaggggaa tgagttggca ctctggaatt gcagacatcg tggatgggga 720 aatcatgact gcagtcacaa tgaggatgtc acattaactt gttatgatag tagtgatctt 780 gaactaaggc ttgtaggtgg aactaaccgc tgtatgggga gagtagagct gaaaatccaa 840 ggaaggtggg ggaccgtatg ccaccataag tggaacaatg ctgcagctga tgtcgtatgc 900 aagcagttgg gatgtggaac cgcacttcac ttcgctggct tgcctcattt gcagtcaggg 960 tctgatgttg tatggcttga tggtgtctcc tgctccggta atgaatcttt tctttgggac 1020 tgcagacatt ccggaaccgt caattttgac tgtcttcatc aaaacgatgt gtctgtgatc 1080 tgctcagatg gagcagattt ggaactgcga ctagcagatg gaagtaacaa ttgttcaggg 1140 agagtagagg tgagaattca tgaacagtgg tggacaatat gtgaccagaa ctggaagaat 1200 gaacaagccc ttgtggtttg taagcagcta ggatgtccgt tcagcgtctt tggcagtcgt 1260 cgtgctaaac ctagtaatga agctagagac atttggataa acagcatatc ttgcactggg 1320 aatgagtcag ctctctggga ctgcacatat gatggaaaag caaagcgaac atgcttccga 1380 agatcagatg ctggagtaat ttgttctgat aaggcagatc tggacctaag gcttgtcggg 1440 gctcatagcc cctgttatgg gagattggag gtgaaatacc aaggagagtg ggggactgtg 1500 tgtcatgaca gatggagcac aaggaatgca gctgttgtgt gtaaacaatt gggatgtgga 1560 aagcctatgc atgtgtttgg tatgacctat tttaaagaag catcaggacc tatttggctg 1620 gatgacgttt cttgcattgg aaatgagtca aatatctggg actgtgaaca cagtggatgg 1680 ggaaagcata attgtgtaca cagagaggat gtgattgtaa cctgctcagg tgatgcaaca 1740 tggggcctga ggctggtggg cggcagcaac cgctgctcgg gaagactgga ggtgtacttt 1800 caaggacggt ggggcacagt gtgtgatgac ggctggaaca gtaaagctgc agctgtggtg 1860 tgtagccagc tggactgccc atcttctatc attggcatgg gtctgggaaa cgcttctaca 1920 ggatatggaa aaatttggct cgatgatgtt tcctgtgatg gagatgagtc agatctctgg 1980 tcatgcagga acagtgggtg gggaaataat gactgcagtc acagtgaaga tgttggagtg 2040 atctgttctg atgcatcgga tatggagctg aggcttgtgg gtggaagcag caggtgtgct 2100 ggaaaagttg aggtgaatgt ccagggtgcc gtgggaattc tgtgtgctaa tggctgggga 2160 atgaacattg ctgaagttgt ttgcaggcaa cttgaatgtg ggtctgcaat cagggtctcc 2220 agagagcctc atttcacaga aagaacatta cacatcttaa tgtcgaattc tggctgcact 2280 ggaggggaag cctctctctg ggattgtata cgatgggagt ggaaacagac tgcgtgtcat 2340 ttaaatatgg aagcaagttt gatctgctca gcccacaggc agcccaggct ggttggagct 2400 gatatgccct gctctggacg tgttgaagtg aaacatgcag acacatggcg ctctgtctgt 2460 gattctgatt tctctcttca tgctgccaat gtgctgtgca gagaattaaa ttgtggagat 2520 gccatatctc tttctgtggg agatcacttt ggaaaaggga atggtctaac ttgggccgaa 2580 aagttccagt gtgaagggag tgaaactcac cttgcattat gccccattgt tcaacatccg 2640 gaagacactt gtatccacag cagagaagtt ggagttgtct gttcccgata tacagatgtc 2700 cgacttgtga atggcaaatc ccagtgtgac gggcaagtgg agatcaacgt gcttggacac 2760 tggggctcac tgtgtgacac ccactgggac ccagaagatg cccgtgttct atgcagacag 2820 ctcagctgtg ggactgctct ctcaaccaca ggaggaaaat atattggaga aagaagtgtt 2880 cgtgtgtggg gacacaggtt tcattgctta gggaatgagt cacttctgga taactgtcaa 2940 atgacagttc ttggagcacc tccctgtatc catggaaata ctgtctctgt gatctgcaca 3000 ggaagcctga cccagccact gtttccatgc ctcgcaaatg tatctgaccc atatttgtct 3060 gcagttccag agggcagtgc tttgatctgc ttagaggaca aacggctccg cctagtggat 3120 ggggacagcc gctgtgccgg gagagtagag atctatcacg acggcttctg gggcaccatc 3180 tgtgatgacg gctgggacct gagcgatgcc cacgtggtgt gtcaaaagct gggctgtgga 3240 gtggccttca atgccacggt ctctgctcac tttggggagg ggtcagggcc catctggctg 3300 gatgacctga actgcacagg aacggagtcc cacttgtggc agtgcccttc ccgcggctgg 3360 gggcagcacg actgcaggca caaggaggac gcaggggtca tctgctcaga attcacagcc 3420 ttgaggctct acagtgaaac tgaaacagag agctgtgctg ggagattgga agtcttctat 3480 aacgggacct ggggcagcgt cggcaggagg aacatcacca cagccatagc aggcattgtg 3540 tgcaggcagc tgggctgtgg ggagaatgga gttgtcagcc tcgccccttt atctaagaca 3600 ggctctggtt tcatgtgggt ggatgacatt cagtgtccta aaacgcatat ctccatatgg 3660 cagtgcctgt ctgccccatg ggagcgaaga atctccagcc cagcagaaga gacctggatc 3720 acatgtgaag atagaataag agtgcgtgga ggagacaccg agtgctctgg gagagtggag 3780 atctggcacg caggctcctg gggcacagtg tgtgatgact cctgggacct ggccgaggcg 3840 gaagtggtgt gtcagcagct gggctgtggc tctgctctgg ctgccctgag ggacgcttcg 3900 tttggccagg gaactggaac catctggttg gatgacatgc ggtgcaaagg aaatgagtca 3960 tttctatggg actgtcacgc caaaccctgg ggacagagtg actgtggaca caaggaagat 4020 gctggcgtga ggtgctctgg acagtcgctg aaatcactga atgcctcctc aggtcattta 4080 gcacttattt tatccagtat ctttgggctc cttctcctgg ttctgtttat tctatttctc 4140 acgtggtgcc gagttcagaa acaaaaacat ctgcccctca gagtttcaac cagaaggagg 4200 ggttctctcg aggagaattt attccatgag atggagacct gcctcaagag agaggaccca 4260 catgggacaa gaacctcaga tgacaccccc aaccatggtt gtgaagatgc tagcgacaca 4320 tcgctgttgg gagttcttcc tgcctctgaa gccacaaaa 4359 11 1453 PRT Homo sapiens 11 Met Met Leu Pro Gln Asn Ser Trp His Ile Asp Phe Gly Arg Cys Cys 1 5 10 15 Cys His Gln Asn Leu Phe Ser Ala Val Val Thr Cys Ile Leu Leu Leu 20 25 30 Asn Ser Cys Phe Leu Ile Ser Ser Phe Asn Gly Thr Asp Leu Glu Leu 35 40 45 Arg Leu Val Asn Gly Asp Gly Pro Cys Ser Gly Thr Val Glu Val Lys 50 55 60 Phe Gln Gly Gln Trp Gly Thr Val Cys Asp Asp Gly Trp Asn Thr Thr 65 70 75 80 Ala Ser Thr Val Val Cys Lys Gln Leu Gly Cys Pro Phe Ser Phe Ala 85 90 95 Met Phe Arg Phe Gly Gln Ala Val Thr Arg His Gly Lys Ile Trp Leu 100 105 110 Asp Asp Val Ser Cys Tyr Gly Asn Glu Ser Ala Leu Trp Glu Cys Gln 115 120 125 His Arg Glu Trp Gly Ser His Asn Cys Tyr His Gly Glu Asp Val Gly 130 135 140 Val Asn Cys Tyr Gly Glu Ala Asn Leu Gly Leu Arg Leu Val Asp Gly 145 150 155 160 Asn Asn Ser Cys Ser Gly Arg Val Glu Val Lys Phe Gln Glu Arg Trp 165 170 175 Gly Thr Ile Cys Asp Asp Gly Trp Asn Leu Asn Thr Ala Ala Val Val 180 185 190 Cys Arg Gln Leu Gly Cys Pro Ser Ser Phe Ile Ser Ser Gly Val Val 195 200 205 Asn Ser Pro Ala Val Leu Arg Pro Ile Trp Leu Asp Asp Ile Leu Cys 210 215 220 Gln Gly Asn Glu Leu Ala Leu Trp Asn Cys Arg His Arg Gly Trp Gly 225 230 235 240 Asn His Asp Cys Ser His Asn Glu Asp Val Thr Leu Thr Cys Tyr Asp 245 250 255 Ser Ser Asp Leu Glu Leu Arg Leu Val Gly Gly Thr Asn Arg Cys Met 260 265 270 Gly Arg Val Glu Leu Lys Ile Gln Gly Arg Trp Gly Thr Val Cys His 275 280 285 His Lys Trp Asn Asn Ala Ala Ala Asp Val Val Cys Lys Gln Leu Gly 290 295 300 Cys Gly Thr Ala Leu His Phe Ala Gly Leu Pro His Leu Gln Ser Gly 305 310 315 320 Ser Asp Val Val Trp Leu Asp Gly Val Ser Cys Ser Gly Asn Glu Ser 325 330 335 Phe Leu Trp Asp Cys Arg His Ser Gly Thr Val Asn Phe Asp Cys Leu 340 345 350 His Gln Asn Asp Val Ser Val Ile Cys Ser Asp Gly Ala Asp Leu Glu 355 360 365 Leu Arg Leu Ala Asp Gly Ser Asn Asn Cys Ser Gly Arg Val Glu Val 370 375 380 Arg Ile His Glu Gln Trp Trp Thr Ile Cys Asp Gln Asn Trp Lys Asn 385 390 395 400 Glu Gln Ala Leu Val Val Cys Lys Gln Leu Gly Cys Pro Phe Ser Val 405 410 415 Phe Gly Ser Arg Arg Ala Lys Pro Ser Asn Glu Ala Arg Asp Ile Trp 420 425 430 Ile Asn Ser Ile Ser Cys Thr Gly Asn Glu Ser Ala Leu Trp Asp Cys 435 440 445 Thr Tyr Asp Gly Lys Ala Lys Arg Thr Cys Phe Arg Arg Ser Asp Ala 450 455 460 Gly Val Ile Cys Ser Asp Lys Ala Asp Leu Asp Leu Arg Leu Val Gly 465 470 475 480 Ala His Ser Pro Cys Tyr Gly Arg Leu Glu Val Lys Tyr Gln Gly Glu 485 490 495 Trp Gly Thr Val Cys His Asp Arg Trp Ser Thr Arg Asn Ala Ala Val 500 505 510 Val Cys Lys Gln Leu Gly Cys Gly Lys Pro Met His Val Phe Gly Met 515 520 525 Thr Tyr Phe Lys Glu Ala Ser Gly Pro Ile Trp Leu Asp Asp Val Ser 530 535 540 Cys Ile Gly Asn Glu Ser Asn Ile Trp Asp Cys Glu His Ser Gly Trp 545 550 555 560 Gly Lys His Asn Cys Val His Arg Glu Asp Val Ile Val Thr Cys Ser 565 570 575 Gly Asp Ala Thr Trp Gly Leu Arg Leu Val Gly Gly Ser Asn Arg Cys 580 585 590 Ser Gly Arg Leu Glu Val Tyr Phe Gln Gly Arg Trp Gly Thr Val Cys 595 600 605 Asp Asp Gly Trp Asn Ser Lys Ala Ala Ala Val Val Cys Ser Gln Leu 610 615 620 Asp Cys Pro Ser Ser Ile Ile Gly Met Gly Leu Gly Asn Ala Ser Thr 625 630 635 640 Gly Tyr Gly Lys Ile Trp Leu Asp Asp Val Ser Cys Asp Gly Asp Glu 645 650 655 Ser Asp Leu Trp Ser Cys Arg Asn Ser Gly Trp Gly Asn Asn Asp Cys 660 665 670 Ser His Ser Glu Asp Val Gly Val Ile Cys Ser Asp Ala Ser Asp Met 675 680 685 Glu Leu Arg Leu Val Gly Gly Ser Ser Arg Cys Ala Gly Lys Val Glu 690 695 700 Val Asn Val Gln Gly Ala Val Gly Ile Leu Cys Ala Asn Gly Trp Gly 705 710 715 720 Met Asn Ile Ala Glu Val Val Cys Arg Gln Leu Glu Cys Gly Ser Ala 725 730 735 Ile Arg Val Ser Arg Glu Pro His Phe Thr Glu Arg Thr Leu His Ile 740 745 750 Leu Met Ser Asn Ser Gly Cys Thr Gly Gly Glu Ala Ser Leu Trp Asp 755 760 765 Cys Ile Arg Trp Glu Trp Lys Gln Thr Ala Cys His Leu Asn Met Glu 770 775 780 Ala Ser Leu Ile Cys Ser Ala His Arg Gln Pro Arg Leu Val Gly Ala 785 790 795 800 Asp Met Pro Cys Ser Gly Arg Val Glu Val Lys His Ala Asp Thr Trp 805 810 815 Arg Ser Val Cys Asp Ser Asp Phe Ser Leu His Ala Ala Asn Val Leu 820 825 830 Cys Arg Glu Leu Asn Cys Gly Asp Ala Ile Ser Leu Ser Val Gly Asp 835 840 845 His Phe Gly Lys Gly Asn Gly Leu Thr Trp Ala Glu Lys Phe Gln Cys 850 855 860 Glu Gly Ser Glu Thr His Leu Ala Leu Cys Pro Ile Val Gln His Pro 865 870 875 880 Glu Asp Thr Cys Ile His Ser Arg Glu Val Gly Val Val Cys Ser Arg 885 890 895 Tyr Thr Asp Val Arg Leu Val Asn Gly Lys Ser Gln Cys Asp Gly Gln 900 905 910 Val Glu Ile Asn Val Leu Gly His Trp Gly Ser Leu Cys Asp Thr His 915 920 925 Trp Asp Pro Glu Asp Ala Arg Val Leu Cys Arg Gln Leu Ser Cys Gly 930 935 940 Thr Ala Leu Ser Thr Thr Gly Gly Lys Tyr Ile Gly Glu Arg Ser Val 945 950 955 960 Arg Val Trp Gly His Arg Phe His Cys Leu Gly Asn Glu Ser Leu Leu 965 970 975 Asp Asn Cys Gln Met Thr Val Leu Gly Ala Pro Pro Cys Ile His Gly 980 985 990 Asn Thr Val Ser Val Ile Cys Thr Gly Ser Leu Thr Gln Pro Leu Phe 995 1000 1005 Pro Cys Leu Ala Asn Val Ser Asp Pro Tyr Leu Ser Ala Val Pro Glu 1010 1015 1020 Gly Ser Ala Leu Ile Cys Leu Glu Asp Lys Arg Leu Arg Leu Val Asp 1025 1030 1035 1040 Gly Asp Ser Arg Cys Ala Gly Arg Val Glu Ile Tyr His Asp Gly Phe 1045 1050 1055 Trp Gly Thr Ile Cys Asp Asp Gly Trp Asp Leu Ser Asp Ala His Val 1060 1065 1070 Val Cys Gln Lys Leu Gly Cys Gly Val Ala Phe Asn Ala Thr Val Ser 1075 1080 1085 Ala His Phe Gly Glu Gly Ser Gly Pro Ile Trp Leu Asp Asp Leu Asn 1090 1095 1100 Cys Thr Gly Thr Glu Ser His Leu Trp Gln Cys Pro Ser Arg Gly Trp 1105 1110 1115 1120 Gly Gln His Asp Cys Arg His Lys Glu Asp Ala Gly Val Ile Cys Ser 1125 1130 1135 Glu Phe Thr Ala Leu Arg Leu Tyr Ser Glu Thr Glu Thr Glu Ser Cys 1140 1145 1150 Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Thr Trp Gly Ser Val Gly 1155 1160 1165 Arg Arg Asn Ile Thr Thr Ala Ile Ala Gly Ile Val Cys Arg Gln Leu 1170 1175 1180 Gly Cys Gly Glu Asn Gly Val Val Ser Leu Ala Pro Leu Ser Lys Thr 1185 1190 1195 1200 Gly Ser Gly Phe Met Trp Val Asp Asp Ile Gln Cys Pro Lys Thr His 1205 1210 1215 Ile Ser Ile Trp Gln Cys Leu Ser Ala Pro Trp Glu Arg Arg Ile Ser 1220 1225 1230 Ser Pro Ala Glu Glu Thr Trp Ile Thr Cys Glu Asp Arg Ile Arg Val 1235 1240 1245 Arg Gly Gly Asp Thr Glu Cys Ser Gly Arg Val Glu Ile Trp His Ala 1250 1255 1260 Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Ala Glu Ala 1265 1270 1275 1280 Glu Val Val Cys Gln Gln Leu Gly Cys Gly Ser Ala Leu Ala Ala Leu 1285 1290 1295 Arg Asp Ala Ser Phe Gly Gln Gly Thr Gly Thr Ile Trp Leu Asp Asp 1300 1305 1310 Met Arg Cys Lys Gly Asn Glu Ser Phe Leu Trp Asp Cys His Ala Lys 1315 1320 1325 Pro Trp Gly Gln Ser Asp Cys Gly His Lys Glu Asp Ala Gly Val Arg 1330 1335 1340 Cys Ser Gly Gln Ser Leu Lys Ser Leu Asn Ala Ser Ser Gly His Leu 1345 1350 1355 1360 Ala Leu Ile Leu Ser Ser Ile Phe Gly Leu Leu Leu Leu Val Leu Phe 1365 1370 1375 Ile Leu Phe Leu Thr Trp Cys Arg Val Gln Lys Gln Lys His Leu Pro 1380 1385 1390 Leu Arg Val Ser Thr Arg Arg Arg Gly Ser Leu Glu Glu Asn Leu Phe 1395 1400 1405 His Glu Met Glu Thr Cys Leu Lys Arg Glu Asp Pro His Gly Thr Arg 1410 1415 1420 Thr Ser Asp Asp Thr Pro Asn His Gly Cys Glu Asp Ala Ser Asp Thr 1425 1430 1435 1440 Ser Leu Leu Gly Val Leu Pro Ala Ser Glu Ala Thr Lys 1445 1450 12 40 PRT Homo sapiens 12 Met Met Leu Pro Gln Asn Ser Trp His Ile Asp Phe Gly Arg Cys Cys 1 5 10 15 Cys His Gln Asn Leu Phe Ser Ala Val Val Thr Cys Ile Leu Leu Leu 20 25 30 Asn Ser Cys Phe Leu Ile Ser Ser 35 40 13 1413 PRT Homo sapiens 13 Phe Asn Gly Thr Asp Leu Glu Leu Arg Leu Val Asn Gly Asp Gly Pro 1 5 10 15 Cys Ser Gly Thr Val Glu Val Lys Phe Gln Gly Gln Trp Gly Thr Val 20 25 30 Cys Asp Asp Gly Trp Asn Thr Thr Ala Ser Thr Val Val Cys Lys Gln 35 40 45 Leu Gly Cys Pro Phe Ser Phe Ala Met Phe Arg Phe Gly Gln Ala Val 50 55 60 Thr Arg His Gly Lys Ile Trp Leu Asp Asp Val Ser Cys Tyr Gly Asn 65 70 75 80 Glu Ser Ala Leu Trp Glu Cys Gln His Arg Glu Trp Gly Ser His Asn 85 90 95 Cys Tyr His Gly Glu Asp Val Gly Val Asn Cys Tyr Gly Glu Ala Asn 100 105 110 Leu Gly Leu Arg Leu Val Asp Gly Asn Asn Ser Cys Ser Gly Arg Val 115 120 125 Glu Val Lys Phe Gln Glu Arg Trp Gly Thr Ile Cys Asp Asp Gly Trp 130 135 140 Asn Leu Asn Thr Ala Ala Val Val Cys Arg Gln Leu Gly Cys Pro Ser 145 150 155 160 Ser Phe Ile Ser Ser Gly Val Val Asn Ser Pro Ala Val Leu Arg Pro 165 170 175 Ile Trp Leu Asp Asp Ile Leu Cys Gln Gly Asn Glu Leu Ala Leu Trp 180 185 190 Asn Cys Arg His Arg Gly Trp Gly Asn His Asp Cys Ser His Asn Glu 195 200 205 Asp Val Thr Leu Thr Cys Tyr Asp Ser Ser Asp Leu Glu Leu Arg Leu 210 215 220 Val Gly Gly Thr Asn Arg Cys Met Gly Arg Val Glu Leu Lys Ile Gln 225 230 235 240 Gly Arg Trp Gly Thr Val Cys His His Lys Trp Asn Asn Ala Ala Ala 245 250 255 Asp Val Val Cys Lys Gln Leu Gly Cys Gly Thr Ala Leu His Phe Ala 260 265 270 Gly Leu Pro His Leu Gln Ser Gly Ser Asp Val Val Trp Leu Asp Gly 275 280 285 Val Ser Cys Ser Gly Asn Glu Ser Phe Leu Trp Asp Cys Arg His Ser 290 295 300 Gly Thr Val Asn Phe Asp Cys Leu His Gln Asn Asp Val Ser Val Ile 305 310 315 320 Cys Ser Asp Gly Ala Asp Leu Glu Leu Arg Leu Ala Asp Gly Ser Asn 325 330 335 Asn Cys Ser Gly Arg Val Glu Val Arg Ile His Glu Gln Trp Trp Thr 340 345 350 Ile Cys Asp Gln Asn Trp Lys Asn Glu Gln Ala Leu Val Val Cys Lys 355 360 365 Gln Leu Gly Cys Pro Phe Ser Val Phe Gly Ser Arg Arg Ala Lys Pro 370 375 380 Ser Asn Glu Ala Arg Asp Ile Trp Ile Asn Ser Ile Ser Cys Thr Gly 385 390 395 400 Asn Glu Ser Ala Leu Trp Asp Cys Thr Tyr Asp Gly Lys Ala Lys Arg 405 410 415 Thr Cys Phe Arg Arg Ser Asp Ala Gly Val Ile Cys Ser Asp Lys Ala 420 425 430 Asp Leu Asp Leu Arg Leu Val Gly Ala His Ser Pro Cys Tyr Gly Arg 435 440 445 Leu Glu Val Lys Tyr Gln Gly Glu Trp Gly Thr Val Cys His Asp Arg 450 455 460 Trp Ser Thr Arg Asn Ala Ala Val Val Cys Lys Gln Leu Gly Cys Gly 465 470 475 480 Lys Pro Met His Val Phe Gly Met Thr Tyr Phe Lys Glu Ala Ser Gly 485 490 495 Pro Ile Trp Leu Asp Asp Val Ser Cys Ile Gly Asn Glu Ser Asn Ile 500 505 510 Trp Asp Cys Glu His Ser Gly Trp Gly Lys His Asn Cys Val His Arg 515 520 525 Glu Asp Val Ile Val Thr Cys Ser Gly Asp Ala Thr Trp Gly Leu Arg 530 535 540 Leu Val Gly Gly Ser Asn Arg Cys Ser Gly Arg Leu Glu Val Tyr Phe 545 550 555 560 Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Gly Trp Asn Ser Lys Ala 565 570 575 Ala Ala Val Val Cys Ser Gln Leu Asp Cys Pro Ser Ser Ile Ile Gly 580 585 590 Met Gly Leu Gly Asn Ala Ser Thr Gly Tyr Gly Lys Ile Trp Leu Asp 595 600 605 Asp Val Ser Cys Asp Gly Asp Glu Ser Asp Leu Trp Ser Cys Arg Asn 610 615 620 Ser Gly Trp Gly Asn Asn Asp Cys Ser His Ser Glu Asp Val Gly Val 625 630 635 640 Ile Cys Ser Asp Ala Ser Asp Met Glu Leu Arg Leu Val Gly Gly Ser 645 650 655 Ser Arg Cys Ala Gly Lys Val Glu Val Asn Val Gln Gly Ala Val Gly 660 665 670 Ile Leu Cys Ala Asn Gly Trp Gly Met Asn Ile Ala Glu Val Val Cys 675 680 685 Arg Gln Leu Glu Cys Gly Ser Ala Ile Arg Val Ser Arg Glu Pro His 690 695 700 Phe Thr Glu Arg Thr Leu His Ile Leu Met Ser Asn Ser Gly Cys Thr 705 710 715 720 Gly Gly Glu Ala Ser Leu Trp Asp Cys Ile Arg Trp Glu Trp Lys Gln 725 730 735 Thr Ala Cys His Leu Asn Met Glu Ala Ser Leu Ile Cys Ser Ala His 740 745 750 Arg Gln Pro Arg Leu Val Gly Ala Asp Met Pro Cys Ser Gly Arg Val 755 760 765 Glu Val Lys His Ala Asp Thr Trp Arg Ser Val Cys Asp Ser Asp Phe 770 775 780 Ser Leu His Ala Ala Asn Val Leu Cys Arg Glu Leu Asn Cys Gly Asp 785 790 795 800 Ala Ile Ser Leu Ser Val Gly Asp His Phe Gly Lys Gly Asn Gly Leu 805 810 815 Thr Trp Ala Glu Lys Phe Gln Cys Glu Gly Ser Glu Thr His Leu Ala 820 825 830 Leu Cys Pro Ile Val Gln His Pro Glu Asp Thr Cys Ile His Ser Arg 835 840 845 Glu Val Gly Val Val Cys Ser Arg Tyr Thr Asp Val Arg Leu Val Asn 850 855 860 Gly Lys Ser Gln Cys Asp Gly Gln Val Glu Ile Asn Val Leu Gly His 865 870 875 880 Trp Gly Ser Leu Cys Asp Thr His Trp Asp Pro Glu Asp Ala Arg Val 885 890 895 Leu Cys Arg Gln Leu Ser Cys Gly Thr Ala Leu Ser Thr Thr Gly Gly 900 905 910 Lys Tyr Ile Gly Glu Arg Ser Val Arg Val Trp Gly His Arg Phe His 915 920 925 Cys Leu Gly Asn Glu Ser Leu Leu Asp Asn Cys Gln Met Thr Val Leu 930 935 940 Gly Ala Pro Pro Cys Ile His Gly Asn Thr Val Ser Val Ile Cys Thr 945 950 955 960 Gly Ser Leu Thr Gln Pro Leu Phe Pro Cys Leu Ala Asn Val Ser Asp 965 970 975 Pro Tyr Leu Ser Ala Val Pro Glu Gly Ser Ala Leu Ile Cys Leu Glu 980 985 990 Asp Lys Arg Leu Arg Leu Val Asp Gly Asp Ser Arg Cys Ala Gly Arg 995 1000 1005 Val Glu Ile Tyr His Asp Gly Phe Trp Gly Thr Ile Cys Asp Asp Gly 1010 1015 1020 Trp Asp Leu Ser Asp Ala His Val Val Cys Gln Lys Leu Gly Cys Gly 1025 1030 1035 1040 Val Ala Phe Asn Ala Thr Val Ser Ala His Phe Gly Glu Gly Ser Gly 1045 1050 1055 Pro Ile Trp Leu Asp Asp Leu Asn Cys Thr Gly Thr Glu Ser His Leu 1060 1065 1070 Trp Gln Cys Pro Ser Arg Gly Trp Gly Gln His Asp Cys Arg His Lys 1075 1080 1085 Glu Asp Ala Gly Val Ile Cys Ser Glu Phe Thr Ala Leu Arg Leu Tyr 1090 1095 1100 Ser Glu Thr Glu Thr Glu Ser Cys Ala Gly Arg Leu Glu Val Phe Tyr 1105 1110 1115 1120 Asn Gly Thr Trp Gly Ser Val Gly Arg Arg Asn Ile Thr Thr Ala Ile 1125 1130 1135 Ala Gly Ile Val Cys Arg Gln Leu Gly Cys Gly Glu Asn Gly Val Val 1140 1145 1150 Ser Leu Ala Pro Leu Ser Lys Thr Gly Ser Gly Phe Met Trp Val Asp 1155 1160 1165 Asp Ile Gln Cys Pro Lys Thr His Ile Ser Ile Trp Gln Cys Leu Ser 1170 1175 1180 Ala Pro Trp Glu Arg Arg Ile Ser Ser Pro Ala Glu Glu Thr Trp Ile 1185 1190 1195 1200 Thr Cys Glu Asp Arg Ile Arg Val Arg Gly Gly Asp Thr Glu Cys Ser 1205 1210 1215 Gly Arg Val Glu Ile Trp His Ala Gly Ser Trp Gly Thr Val Cys Asp 1220 1225 1230 Asp Ser Trp Asp Leu Ala Glu Ala Glu Val Val Cys Gln Gln Leu Gly 1235 1240 1245 Cys Gly Ser Ala Leu Ala Ala Leu Arg Asp Ala Ser Phe Gly Gln Gly 1250 1255 1260 Thr Gly Thr Ile Trp Leu Asp Asp Met Arg Cys Lys Gly Asn Glu Ser 1265 1270 1275 1280 Phe Leu Trp Asp Cys His Ala Lys Pro Trp Gly Gln Ser Asp Cys Gly 1285 1290 1295 His Lys Glu Asp Ala Gly Val Arg Cys Ser Gly Gln Ser Leu Lys Ser 1300 1305 1310 Leu Asn Ala Ser Ser Gly His Leu Ala Leu Ile Leu Ser Ser Ile Phe 1315 1320 1325 Gly Leu Leu Leu Leu Val Leu Phe Ile Leu Phe Leu Thr Trp Cys Arg 1330 1335 1340 Val Gln Lys Gln Lys His Leu Pro Leu Arg Val Ser Thr Arg Arg Arg 1345 1350 1355 1360 Gly Ser Leu Glu Glu Asn Leu Phe His Glu Met Glu Thr Cys Leu Lys 1365 1370 1375 Arg Glu Asp Pro His Gly Thr Arg Thr Ser Asp Asp Thr Pro Asn His 1380 1385 1390 Gly Cys Glu Asp Ala Ser Asp Thr Ser Leu Leu Gly Val Leu Pro Ala 1395 1400 1405 Ser Glu Ala Thr Lys 1410 14 1319 PRT Homo sapiens 14 Phe Asn Gly Thr Asp Leu Glu Leu Arg Leu Val Asn Gly Asp Gly Pro 1 5 10 15 Cys Ser Gly Thr Val Glu Val Lys Phe Gln Gly Gln Trp Gly Thr Val 20 25 30 Cys Asp Asp Gly Trp Asn Thr Thr Ala Ser Thr Val Val Cys Lys Gln 35 40 45 Leu Gly Cys Pro Phe Ser Phe Ala Met Phe Arg Phe Gly Gln Ala Val 50 55 60 Thr Arg His Gly Lys Ile Trp Leu Asp Asp Val Ser Cys Tyr Gly Asn 65 70 75 80 Glu Ser Ala Leu Trp Glu Cys Gln His Arg Glu Trp Gly Ser His Asn 85 90 95 Cys Tyr His Gly Glu Asp Val Gly Val Asn Cys Tyr Gly Glu Ala Asn 100 105 110 Leu Gly Leu Arg Leu Val Asp Gly Asn Asn Ser Cys Ser Gly Arg Val 115 120 125 Glu Val Lys Phe Gln Glu Arg Trp Gly Thr Ile Cys Asp Asp Gly Trp 130 135 140 Asn Leu Asn Thr Ala Ala Val Val Cys Arg Gln Leu Gly Cys Pro Ser 145 150 155 160 Ser Phe Ile Ser Ser Gly Val Val Asn Ser Pro Ala Val Leu Arg Pro 165 170 175 Ile Trp Leu Asp Asp Ile Leu Cys Gln Gly Asn Glu Leu Ala Leu Trp 180 185 190 Asn Cys Arg His Arg Gly Trp Gly Asn His Asp Cys Ser His Asn Glu 195 200 205 Asp Val Thr Leu Thr Cys Tyr Asp Ser Ser Asp Leu Glu Leu Arg Leu 210 215 220 Val Gly Gly Thr Asn Arg Cys Met Gly Arg Val Glu Leu Lys Ile Gln 225 230 235 240 Gly Arg Trp Gly Thr Val Cys His His Lys Trp Asn Asn Ala Ala Ala 245 250 255 Asp Val Val Cys Lys Gln Leu Gly Cys Gly Thr Ala Leu His Phe Ala 260 265 270 Gly Leu Pro His Leu Gln Ser Gly Ser Asp Val Val Trp Leu Asp Gly 275 280 285 Val Ser Cys Ser Gly Asn Glu Ser Phe Leu Trp Asp Cys Arg His Ser 290 295 300 Gly Thr Val Asn Phe Asp Cys Leu His Gln Asn Asp Val Ser Val Ile 305 310 315 320 Cys Ser Asp Gly Ala Asp Leu Glu Leu Arg Leu Ala Asp Gly Ser Asn 325 330 335 Asn Cys Ser Gly Arg Val Glu Val Arg Ile His Glu Gln Trp Trp Thr 340 345 350 Ile Cys Asp Gln Asn Trp Lys Asn Glu Gln Ala Leu Val Val Cys Lys 355 360 365 Gln Leu Gly Cys Pro Phe Ser Val Phe Gly Ser Arg Arg Ala Lys Pro 370 375 380 Ser Asn Glu Ala Arg Asp Ile Trp Ile Asn Ser Ile Ser Cys Thr Gly 385 390 395 400 Asn Glu Ser Ala Leu Trp Asp Cys Thr Tyr Asp Gly Lys Ala Lys Arg 405 410 415 Thr Cys Phe Arg Arg Ser Asp Ala Gly Val Ile Cys Ser Asp Lys Ala 420 425 430 Asp Leu Asp Leu Arg Leu Val Gly Ala His Ser Pro Cys Tyr Gly Arg 435 440 445 Leu Glu Val Lys Tyr Gln Gly Glu Trp Gly Thr Val Cys His Asp Arg 450 455 460 Trp Ser Thr Arg Asn Ala Ala Val Val Cys Lys Gln Leu Gly Cys Gly 465 470 475 480 Lys Pro Met His Val Phe Gly Met Thr Tyr Phe Lys Glu Ala Ser Gly 485 490 495 Pro Ile Trp Leu Asp Asp Val Ser Cys Ile Gly Asn Glu Ser Asn Ile 500 505 510 Trp Asp Cys Glu His Ser Gly Trp Gly Lys His Asn Cys Val His Arg 515 520 525 Glu Asp Val Ile Val Thr Cys Ser Gly Asp Ala Thr Trp Gly Leu Arg 530 535 540 Leu Val Gly Gly Ser Asn Arg Cys Ser Gly Arg Leu Glu Val Tyr Phe 545 550 555 560 Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Gly Trp Asn Ser Lys Ala 565 570 575 Ala Ala Val Val Cys Ser Gln Leu Asp Cys Pro Ser Ser Ile Ile Gly 580 585 590 Met Gly Leu Gly Asn Ala Ser Thr Gly Tyr Gly Lys Ile Trp Leu Asp 595 600 605 Asp Val Ser Cys Asp Gly Asp Glu Ser Asp Leu Trp Ser Cys Arg Asn 610 615 620 Ser Gly Trp Gly Asn Asn Asp Cys Ser His Ser Glu Asp Val Gly Val 625 630 635 640 Ile Cys Ser Asp Ala Ser Asp Met Glu Leu Arg Leu Val Gly Gly Ser 645 650 655 Ser Arg Cys Ala Gly Lys Val Glu Val Asn Val Gln Gly Ala Val Gly 660 665 670 Ile Leu Cys Ala Asn Gly Trp Gly Met Asn Ile Ala Glu Val Val Cys 675 680 685 Arg Gln Leu Glu Cys Gly Ser Ala Ile Arg Val Ser Arg Glu Pro His 690 695 700 Phe Thr Glu Arg Thr Leu His Ile Leu Met Ser Asn Ser Gly Cys Thr 705 710 715 720 Gly Gly Glu Ala Ser Leu Trp Asp Cys Ile Arg Trp Glu Trp Lys Gln 725 730 735 Thr Ala Cys His Leu Asn Met Glu Ala Ser Leu Ile Cys Ser Ala His 740 745 750 Arg Gln Pro Arg Leu Val Gly Ala Asp Met Pro Cys Ser Gly Arg Val 755 760 765 Glu Val Lys His Ala Asp Thr Trp Arg Ser Val Cys Asp Ser Asp Phe 770 775 780 Ser Leu His Ala Ala Asn Val Leu Cys Arg Glu Leu Asn Cys Gly Asp 785 790 795 800 Ala Ile Ser Leu Ser Val Gly Asp His Phe Gly Lys Gly Asn Gly Leu 805 810 815 Thr Trp Ala Glu Lys Phe Gln Cys Glu Gly Ser Glu Thr His Leu Ala 820 825 830 Leu Cys Pro Ile Val Gln His Pro Glu Asp Thr Cys Ile His Ser Arg 835 840 845 Glu Val Gly Val Val Cys Ser Arg Tyr Thr Asp Val Arg Leu Val Asn 850 855 860 Gly Lys Ser Gln Cys Asp Gly Gln Val Glu Ile Asn Val Leu Gly His 865 870 875 880 Trp Gly Ser Leu Cys Asp Thr His Trp Asp Pro Glu Asp Ala Arg Val 885 890 895 Leu Cys Arg Gln Leu Ser Cys Gly Thr Ala Leu Ser Thr Thr Gly Gly 900 905 910 Lys Tyr Ile Gly Glu Arg Ser Val Arg Val Trp Gly His Arg Phe His 915 920 925 Cys Leu Gly Asn Glu Ser Leu Leu Asp Asn Cys Gln Met Thr Val Leu 930 935 940 Gly Ala Pro Pro Cys Ile His Gly Asn Thr Val Ser Val Ile Cys Thr 945 950 955 960 Gly Ser Leu Thr Gln Pro Leu Phe Pro Cys Leu Ala Asn Val Ser Asp 965 970 975 Pro Tyr Leu Ser Ala Val Pro Glu Gly Ser Ala Leu Ile Cys Leu Glu 980 985 990 Asp Lys Arg Leu Arg Leu Val Asp Gly Asp Ser Arg Cys Ala Gly Arg 995 1000 1005 Val Glu Ile Tyr His Asp Gly Phe Trp Gly Thr Ile Cys Asp Asp Gly 1010 1015 1020 Trp Asp Leu Ser Asp Ala His Val Val Cys Gln Lys Leu Gly Cys Gly 1025 1030 1035 1040 Val Ala Phe Asn Ala Thr Val Ser Ala His Phe Gly Glu Gly Ser Gly 1045 1050 1055 Pro Ile Trp Leu Asp Asp Leu Asn Cys Thr Gly Thr Glu Ser His Leu 1060 1065 1070 Trp Gln Cys Pro Ser Arg Gly Trp Gly Gln His Asp Cys Arg His Lys 1075 1080 1085 Glu Asp Ala Gly Val Ile Cys Ser Glu Phe Thr Ala Leu Arg Leu Tyr 1090 1095 1100 Ser Glu Thr Glu Thr Glu Ser Cys Ala Gly Arg Leu Glu Val Phe Tyr 1105 1110 1115 1120 Asn Gly Thr Trp Gly Ser Val Gly Arg Arg Asn Ile Thr Thr Ala Ile 1125 1130 1135 Ala Gly Ile Val Cys Arg Gln Leu Gly Cys Gly Glu Asn Gly Val Val 1140 1145 1150 Ser Leu Ala Pro Leu Ser Lys Thr Gly Ser Gly Phe Met Trp Val Asp 1155 1160 1165 Asp Ile Gln Cys Pro Lys Thr His Ile Ser Ile Trp Gln Cys Leu Ser 1170 1175 1180 Ala Pro Trp Glu Arg Arg Ile Ser Ser Pro Ala Glu Glu Thr Trp Ile 1185 1190 1195 1200 Thr Cys Glu Asp Arg Ile Arg Val Arg Gly Gly Asp Thr Glu Cys Ser 1205 1210 1215 Gly Arg Val Glu Ile Trp His Ala Gly Ser Trp Gly Thr Val Cys Asp 1220 1225 1230 Asp Ser Trp Asp Leu Ala Glu Ala Glu Val Val Cys Gln Gln Leu Gly 1235 1240 1245 Cys Gly Ser Ala Leu Ala Ala Leu Arg Asp Ala Ser Phe Gly Gln Gly 1250 1255 1260 Thr Gly Thr Ile Trp Leu Asp Asp Met Arg Cys Lys Gly Asn Glu Ser 1265 1270 1275 1280 Phe Leu Trp Asp Cys His Ala Lys Pro Trp Gly Gln Ser Asp Cys Gly 1285 1290 1295 His Lys Glu Asp Ala Gly Val Arg Cys Ser Gly Gln Ser Leu Lys Ser 1300 1305 1310 Leu Asn Ala Ser Ser Gly His 1315 15 24 PRT Homo sapiens 15 Leu Ala Leu Ile Leu Ser Ser Ile Phe Gly Leu Leu Leu Leu Val Leu 1 5 10 15 Phe Ile Leu Phe Leu Thr Trp Cys 20 16 70 PRT Homo sapiens 16 Arg Val Gln Lys Gln Lys His Leu Pro Leu Arg Val Ser Thr Arg Arg 1 5 10 15 Arg Gly Ser Leu Glu Glu Asn Leu Phe His Glu Met Glu Thr Cys Leu 20 25 30 Lys Arg Glu Asp Pro His Gly Thr Arg Thr Ser Asp Asp Thr Pro Asn 35 40 45 His Gly Cys Glu Asp Ala Ser Asp Thr Ser Leu Leu Gly Val Leu Pro 50 55 60 Ala Ser Glu Ala Thr Lys 65 70 17 3104 DNA Homo sapiens 17 gtcgacccac gcgtccggtc tgtggctgag catggccctc ccagccctgg gcctggaccc 60 ctggagcctc ctgggccttt tcctcttcca actgcttcag ctgctgctgc cgacgacgac 120 cgcgggggga ggcgggcagg ggcccatgcc cagggtcaga tactatgcag gggatgaacg 180 tagggcactt agcttcttcc accagaaggg cctccaggat tttgacactc tgctcctgag 240 tggtgatgga aatactctct acgtgggggc tcgagaagcc attctggcct tggatatcca 300 ggatccaggg gtccccaggc taaagaacat gataccgtgg ccagccagtg acagaaaaaa 360 gagtgaatgt gcctttaaga agaagagcaa tgagacacag tgtttcaact tcatccgtgt 420 cctggtttct tacaatgtca cccatctcta cacctgcggc accttcgcct tcagccctgc 480 ttgtaccttc attgaacttc aagattccta cctgttgccc atctcggagg acaaggtcat 540 ggagggaaaa ggccaaagcc cctttgaccc cgctcacaag catacggctg tcttggtgga 600 tgggatgctc tattctggta ctatgaacaa cttcctgggc agtgagccca tcctgatgcg 660 cacactggga tcccagcctg tcctcaagac cgacaacttc ctccgctggc tgcatcatga 720 cgcctccttt gtggcagcca tcccttcgac ccaggtcgtc tacttcttct tcgaggagac 780 agccagcgag tttgacttct ttgagaggct ccacacatcg cgggtggcta gagtctgcaa 840 gaatgacgtg ggcggcgaaa agctgctgca gaagaagtgg accaccttcc tgaaggccca 900 gctgctctgc acccagccgg ggcagctgcc cttcaacgtc atccgccacg cggtcctgct 960 ccccgccgat tctcccacag ctccccacat ctacgcagtc ttcacctccc agtggcaggt 1020 tggcgggacc aggagctctg cggtttgtgc cttctctctc ttggacattg aacgtgtctt 1080 taaggggaaa tacaaagagt tgaacaaaga aacttcacgc tggactactt ataggggccc 1140 tgagaccaac ccccggccag gcagttgctc agtgggcccc tcctctgata aggccctgac 1200 cttcatgaag gaccatttcc tgatggatga gcaagtggtg gggacgcccc tgctggtgaa 1260 atctggcgtg gagtatacac ggcttgcagt ggagacagcc cagggccttg atgggcacag 1320 ccatcttgtc atgtacctgg gaaccaccac agggtcgctc cacaaggctg tggtaagtgg 1380 ggacagcagt gctcatctgg tggaagagat tcagctgttc cctgaccctg aacctgttcg 1440 caacctgcag ctggccccca cccagggtgc agtgtttgta ggcttctcag gaggtgtctg 1500 gagggtgccc cgagccaact gtagtgtcta tgagagctgt gtggactgtg tccttgcccg 1560 ggacccccac tgtgcctggg accctgagtc ccgaacctgt tgcctcctgt ctgcccccaa 1620 cctgaactcc tggaagcagg acatggagcg ggggaaccca gagtgggcat gtgccagtgg 1680 ccccatgagc aggagccttc ggcctcagag ccgcccgcaa atcattaaag aagtcctggc 1740 tgtccccaac tccatcctgg agctcccctg cccccacctg tcagccttgg cctcttatta 1800 ttggagtcat ggcccagcag cagtcccaga agcctcttcc actgtctaca atggctccct 1860 cttgctgata gtgcaggatg gagttggggg tctctaccag tgctgggcaa ctgagaatgg 1920 cttttcatac cctgtgatct cctactgggt ggacagccag gaccagaccc tggccctgga 1980 tcctgaactg gcaggcatcc cccgggagca tgtgaaggtc ccgttgacca gggtcagtgg 2040 tggggccgcc ctggctgccc agcagtccta ctggccccac tttgtcactg tcactgtcct 2100 ctttgcctta gtgctttcag gagccctcat catcctcgtg gcctccccat tgagagcact 2160 ccgggctcgg ggcaaggttc agggctgtga gaccctgcgc cctggggaga aggccccgtt 2220 aagcagagag caacacctcc agtctcccaa ggaatgcagg acctctgcca gtgatgtgga 2280 cgctgacaac aactgcctag gcactgaggt agcttaaact ctaggcacag gccggggctg 2340 cggtgcaggc acctggccat gctggctggg cggcccaagc acagccctga ctaggatgac 2400 agcagcacaa aagaccacct ttctcccctg agaggagctt ctgctactct gcatcactga 2460 tgacactcag cagggtgatg cacagcagtc tgcctcccct atgggactcc cttctaccaa 2520 gcacatgagc tctctaacag ggtgggggct acccccagac ctgctcctac actgatattg 2580 aagaacctgg agaggatcct tcagttctgg ccattccagg gaccctccag aaacacagtg 2640 tttcaagaga tcctaaaaaa acctgcctgt cccaggaccc tatggtaatg aacaccaaac 2700 atctaaacaa tcatatgcta acatgccact cctggaaact ccactctgaa gctgccgctt 2760 tggacaccaa cactcccttc tcccagggtc atgcagggat ctgctccctc ctgcttccct 2820 taccagtcgt gcaccgctga ctcccaggaa gtctttcctg aagtctgacc acctttcttc 2880 ttgcttcagt tggggcagac tctgatccct tctgccctgg cagaatggca ggggtaatct 2940 gagccttctt cactccttta ccctagctga ccccttcacc tctccccctc ccttttcctt 3000 tgttttggga ttcagaaaac tgcttgtcag agactgttta ttttttatta aaaatataag 3060 gcttaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagggcgg ccgc 3104 18 2283 DNA Homo sapiens 18 atggccctcc cagccctggg cctggacccc tggagcctcc tgggcctttt cctcttccaa 60 ctgcttcagc tgctgctgcc gacgacgacc gcggggggag gcgggcaggg gcccatgccc 120 agggtcagat actatgcagg ggatgaacgt agggcactta gcttcttcca ccagaagggc 180 ctccaggatt ttgacactct gctcctgagt ggtgatggaa atactctcta cgtgggggct 240 cgagaagcca ttctggcctt ggatatccag gatccagggg tccccaggct aaagaacatg 300 ataccgtggc cagccagtga cagaaaaaag agtgaatgtg cctttaagaa gaagagcaat 360 gagacacagt gtttcaactt catccgtgtc ctggtttctt acaatgtcac ccatctctac 420 acctgcggca ccttcgcctt cagccctgct tgtaccttca ttgaacttca agattcctac 480 ctgttgccca tctcggagga caaggtcatg gagggaaaag gccaaagccc ctttgacccc 540 gctcacaagc atacggctgt cttggtggat gggatgctct attctggtac tatgaacaac 600 ttcctgggca gtgagcccat cctgatgcgc acactgggat cccagcctgt cctcaagacc 660 gacaacttcc tccgctggct gcatcatgac gcctcctttg tggcagccat cccttcgacc 720 caggtcgtct acttcttctt cgaggagaca gccagcgagt ttgacttctt tgagaggctc 780 cacacatcgc gggtggctag agtctgcaag aatgacgtgg gcggcgaaaa gctgctgcag 840 aagaagtgga ccaccttcct gaaggcccag ctgctctgca cccagccggg gcagctgccc 900 ttcaacgtca tccgccacgc ggtcctgctc cccgccgatt ctcccacagc tccccacatc 960 tacgcagtct tcacctccca gtggcaggtt ggcgggacca ggagctctgc ggtttgtgcc 1020 ttctctctct tggacattga acgtgtcttt aaggggaaat acaaagagtt gaacaaagaa 1080 acttcacgct ggactactta taggggccct gagaccaacc cccggccagg cagttgctca 1140 gtgggcccct cctctgataa ggccctgacc ttcatgaagg accatttcct gatggatgag 1200 caagtggtgg ggacgcccct gctggtgaaa tctggcgtgg agtatacacg gcttgcagtg 1260 gagacagccc agggccttga tgggcacagc catcttgtca tgtacctggg aaccaccaca 1320 gggtcgctcc acaaggctgt ggtaagtggg gacagcagtg ctcatctggt ggaagagatt 1380 cagctgttcc ctgaccctga acctgttcgc aacctgcagc tggcccccac ccagggtgca 1440 gtgtttgtag gcttctcagg aggtgtctgg agggtgcccc gagccaactg tagtgtctat 1500 gagagctgtg tggactgtgt ccttgcccgg gacccccact gtgcctggga ccctgagtcc 1560 cgaacctgtt gcctcctgtc tgcccccaac ctgaactcct ggaagcagga catggagcgg 1620 gggaacccag agtgggcatg tgccagtggc cccatgagca ggagccttcg gcctcagagc 1680 cgcccgcaaa tcattaaaga agtcctggct gtccccaact ccatcctgga gctcccctgc 1740 ccccacctgt cagccttggc ctcttattat tggagtcatg gcccagcagc agtcccagaa 1800 gcctcttcca ctgtctacaa tggctccctc ttgctgatag tgcaggatgg agttgggggt 1860 ctctaccagt gctgggcaac tgagaatggc ttttcatacc ctgtgatctc ctactgggtg 1920 gacagccagg accagaccct ggccctggat cctgaactgg caggcatccc ccgggagcat 1980 gtgaaggtcc cgttgaccag ggtcagtggt ggggccgccc tggctgccca gcagtcctac 2040 tggccccact ttgtcactgt cactgtcctc tttgccttag tgctttcagg agccctcatc 2100 atcctcgtgg cctccccatt gagagcactc cgggctcggg gcaaggttca gggctgtgag 2160 accctgcgcc ctggggagaa ggccccgtta agcagagagc aacacctcca gtctcccaag 2220 gaatgcagga cctctgccag tgatgtggac gctgacaaca actgcctagg cactgaggta 2280 gct 2283 19 761 PRT Homo sapiens 19 Met Ala Leu Pro Ala Leu Gly Leu Asp Pro Trp Ser Leu Leu Gly Leu 1 5 10 15 Phe Leu Phe Gln Leu Leu Gln Leu Leu Leu Pro Thr Thr Thr Ala Gly 20 25 30 Gly Gly Gly Gln Gly Pro Met Pro Arg Val Arg Tyr Tyr Ala Gly Asp 35 40 45 Glu Arg Arg Ala Leu Ser Phe Phe His Gln Lys Gly Leu Gln Asp Phe 50 55 60 Asp Thr Leu Leu Leu Ser Gly Asp Gly Asn Thr Leu Tyr Val Gly Ala 65 70 75 80 Arg Glu Ala Ile Leu Ala Leu Asp Ile Gln Asp Pro Gly Val Pro Arg 85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Asp Arg Lys Lys Ser Glu 100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn Phe Ile 115 120 125 Arg Val Leu Val Ser Tyr Asn Val Thr His Leu Tyr Thr Cys Gly Thr 130 135 140 Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser Tyr 145 150 155 160 Leu Leu Pro Ile Ser Glu Asp Lys Val Met Glu Gly Lys Gly Gln Ser 165 170 175 Pro Phe Asp Pro Ala His Lys His Thr Ala Val Leu Val Asp Gly Met 180 185 190 Leu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile Leu 195 200 205 Met Arg Thr Leu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe Leu 210 215 220 Arg Trp Leu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser Thr 225 230 235 240 Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245 250 255 Phe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys Lys Asn Asp 260 265 270 Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu Lys 275 280 285 Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln Leu Pro Phe Asn Val Ile 290 295 300 Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Thr Ala Pro His Ile 305 310 315 320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser Ser 325 330 335 Ala Val Cys Ala Phe Ser Leu Leu Asp Ile Glu Arg Val Phe Lys Gly 340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr Tyr Arg 355 360 365 Gly Pro Glu Thr Asn Pro Arg Pro Gly Ser Cys Ser Val Gly Pro Ser 370 375 380 Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe Leu Met Asp Glu 385 390 395 400 Gln Val Val Gly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr Thr 405 410 415 Arg Leu Ala Val Glu Thr Ala Gln Gly Leu Asp Gly His Ser His Leu 420 425 430 Val Met Tyr Leu Gly Thr Thr Thr Gly Ser Leu His Lys Ala Val Val 435 440 445 Ser Gly Asp Ser Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe Pro 450 455 460 Asp Pro Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly Ala 465 470 475 480 Val Phe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala Asn 485 490 495 Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro 500 505 510 His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys Cys Leu Leu Ser Ala 515 520 525 Pro Asn Leu Asn Ser Trp Lys Gln Asp Met Glu Arg Gly Asn Pro Glu 530 535 540 Trp Ala Cys Ala Ser Gly Pro Met Ser Arg Ser Leu Arg Pro Gln Ser 545 550 555 560 Arg Pro Gln Ile Ile Lys Glu Val Leu Ala Val Pro Asn Ser Ile Leu 565 570 575 Glu Leu Pro Cys Pro His Leu Ser Ala Leu Ala Ser Tyr Tyr Trp Ser 580 585 590 His Gly Pro Ala Ala Val Pro Glu Ala Ser Ser Thr Val Tyr Asn Gly 595 600 605 Ser Leu Leu Leu Ile Val Gln Asp Gly Val Gly Gly Leu Tyr Gln Cys 610 615 620 Trp Ala Thr Glu Asn Gly Phe Ser Tyr Pro Val Ile Ser Tyr Trp Val 625 630 635 640 Asp Ser Gln Asp Gln Thr Leu Ala Leu Asp Pro Glu Leu Ala Gly Ile 645 650 655 Pro Arg Glu His Val Lys Val Pro Leu Thr Arg Val Ser Gly Gly Ala 660 665 670 Ala Leu Ala Ala Gln Gln Ser Tyr Trp Pro His Phe Val Thr Val Thr 675 680 685 Val Leu Phe Ala Leu Val Leu Ser Gly Ala Leu Ile Ile Leu Val Ala 690 695 700 Ser Pro Leu Arg Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Glu 705 710 715 720 Thr Leu Arg Pro Gly Glu Lys Ala Pro Leu Ser Arg Glu Gln His Leu 725 730 735 Gln Ser Pro Lys Glu Cys Arg Thr Ser Ala Ser Asp Val Asp Ala Asp 740 745 750 Asn Asn Cys Leu Gly Thr Glu Val Ala 755 760 20 31 PRT Homo sapiens 20 Met Ala Leu Pro Ala Leu Gly Leu Asp Pro Trp Ser Leu Leu Gly Leu 1 5 10 15 Phe Leu Phe Gln Leu Leu Gln Leu Leu Leu Pro Thr Thr Thr Ala 20 25 30 21 730 PRT Homo sapiens 21 Gly Gly Gly Gly Gln Gly Pro Met Pro Arg Val Arg Tyr Tyr Ala Gly 1 5 10 15 Asp Glu Arg Arg Ala Leu Ser Phe Phe His Gln Lys Gly Leu Gln Asp 20 25 30 Phe Asp Thr Leu Leu Leu Ser Gly Asp Gly Asn Thr Leu Tyr Val Gly 35 40 45 Ala Arg Glu Ala Ile Leu Ala Leu Asp Ile Gln Asp Pro Gly Val Pro 50 55 60 Arg Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Asp Arg Lys Lys Ser 65 70 75 80 Glu Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn Phe 85 90 95 Ile Arg Val Leu Val Ser Tyr Asn Val Thr His Leu Tyr Thr Cys Gly 100 105 110 Thr Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser 115 120 125 Tyr Leu Leu Pro Ile Ser Glu Asp Lys Val Met Glu Gly Lys Gly Gln 130 135 140 Ser Pro Phe Asp Pro Ala His Lys His Thr Ala Val Leu Val Asp Gly 145 150 155 160 Met Leu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile 165 170 175 Leu Met Arg Thr Leu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe 180 185 190 Leu Arg Trp Leu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser 195 200 205 Thr Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp 210 215 220 Phe Phe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys Lys Asn 225 230 235 240 Asp Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu 245 250 255 Lys Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln Leu Pro Phe Asn Val 260 265 270 Ile Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Thr Ala Pro His 275 280 285 Ile Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser 290 295 300 Ser Ala Val Cys Ala Phe Ser Leu Leu Asp Ile Glu Arg Val Phe Lys 305 310 315 320 Gly Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr Tyr 325 330 335 Arg Gly Pro Glu Thr Asn Pro Arg Pro Gly Ser Cys Ser Val Gly Pro 340 345 350 Ser Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe Leu Met Asp 355 360 365 Glu Gln Val Val Gly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr 370 375 380 Thr Arg Leu Ala Val Glu Thr Ala Gln Gly Leu Asp Gly His Ser His 385 390 395 400 Leu Val Met Tyr Leu Gly Thr Thr Thr Gly Ser Leu His Lys Ala Val 405 410 415 Val Ser Gly Asp Ser Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe 420 425 430 Pro Asp Pro Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly 435 440 445 Ala Val Phe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala 450 455 460 Asn Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp 465 470 475 480 Pro His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys Cys Leu Leu Ser 485 490 495 Ala Pro Asn Leu Asn Ser Trp Lys Gln Asp Met Glu Arg Gly Asn Pro 500 505 510 Glu Trp Ala Cys Ala Ser Gly Pro Met Ser Arg Ser Leu Arg Pro Gln 515 520 525 Ser Arg Pro Gln Ile Ile Lys Glu Val Leu Ala Val Pro Asn Ser Ile 530 535 540 Leu Glu Leu Pro Cys Pro His Leu Ser Ala Leu Ala Ser Tyr Tyr Trp 545 550 555 560 Ser His Gly Pro Ala Ala Val Pro Glu Ala Ser Ser Thr Val Tyr Asn 565 570 575 Gly Ser Leu Leu Leu Ile Val Gln Asp Gly Val Gly Gly Leu Tyr Gln 580 585 590 Cys Trp Ala Thr Glu Asn Gly Phe Ser Tyr Pro Val Ile Ser Tyr Trp 595 600 605 Val Asp Ser Gln Asp Gln Thr Leu Ala Leu Asp Pro Glu Leu Ala Gly 610 615 620 Ile Pro Arg Glu His Val Lys Val Pro Leu Thr Arg Val Ser Gly Gly 625 630 635 640 Ala Ala Leu Ala Ala Gln Gln Ser Tyr Trp Pro His Phe Val Thr Val 645 650 655 Thr Val Leu Phe Ala Leu Val Leu Ser Gly Ala Leu Ile Ile Leu Val 660 665 670 Ala Ser Pro Leu Arg Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys 675 680 685 Glu Thr Leu Arg Pro Gly Glu Lys Ala Pro Leu Ser Arg Glu Gln His 690 695 700 Leu Gln Ser Pro Lys Glu Cys Arg Thr Ser Ala Ser Asp Val Asp Ala 705 710 715 720 Asp Asn Asn Cys Leu Gly Thr Glu Val Ala 725 730 22 652 PRT Homo sapiens 22 Gly Gly Gly Gly Gln Gly Pro Met Pro Arg Val Arg Tyr Tyr Ala Gly 1 5 10 15 Asp Glu Arg Arg Ala Leu Ser Phe Phe His Gln Lys Gly Leu Gln Asp 20 25 30 Phe Asp Thr Leu Leu Leu Ser Gly Asp Gly Asn Thr Leu Tyr Val Gly 35 40 45 Ala Arg Glu Ala Ile Leu Ala Leu Asp Ile Gln Asp Pro Gly Val Pro 50 55 60 Arg Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Asp Arg Lys Lys Ser 65 70 75 80 Glu Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn Phe 85 90 95 Ile Arg Val Leu Val Ser Tyr Asn Val Thr His Leu Tyr Thr Cys Gly 100 105 110 Thr Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser 115 120 125 Tyr Leu Leu Pro Ile Ser Glu Asp Lys Val Met Glu Gly Lys Gly Gln 130 135 140 Ser Pro Phe Asp Pro Ala His Lys His Thr Ala Val Leu Val Asp Gly 145 150 155 160 Met Leu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile 165 170 175 Leu Met Arg Thr Leu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe 180 185 190 Leu Arg Trp Leu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser 195 200 205 Thr Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp 210 215 220 Phe Phe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys Lys Asn 225 230 235 240 Asp Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu 245 250 255 Lys Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln Leu Pro Phe Asn Val 260 265 270 Ile Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Thr Ala Pro His 275 280 285 Ile Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser 290 295 300 Ser Ala Val Cys Ala Phe Ser Leu Leu Asp Ile Glu Arg Val Phe Lys 305 310 315 320 Gly Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr Tyr 325 330 335 Arg Gly Pro Glu Thr Asn Pro Arg Pro Gly Ser Cys Ser Val Gly Pro 340 345 350 Ser Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe Leu Met Asp 355 360 365 Glu Gln Val Val Gly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr 370 375 380 Thr Arg Leu Ala Val Glu Thr Ala Gln Gly Leu Asp Gly His Ser His 385 390 395 400 Leu Val Met Tyr Leu Gly Thr Thr Thr Gly Ser Leu His Lys Ala Val 405 410 415 Val Ser Gly Asp Ser Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe 420 425 430 Pro Asp Pro Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly 435 440 445 Ala Val Phe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala 450 455 460 Asn Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp 465 470 475 480 Pro His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys Cys Leu Leu Ser 485 490 495 Ala Pro Asn Leu Asn Ser Trp Lys Gln Asp Met Glu Arg Gly Asn Pro 500 505 510 Glu Trp Ala Cys Ala Ser Gly Pro Met Ser Arg Ser Leu Arg Pro Gln 515 520 525 Ser Arg Pro Gln Ile Ile Lys Glu Val Leu Ala Val Pro Asn Ser Ile 530 535 540 Leu Glu Leu Pro Cys Pro His Leu Ser Ala Leu Ala Ser Tyr Tyr Trp 545 550 555 560 Ser His Gly Pro Ala Ala Val Pro Glu Ala Ser Ser Thr Val Tyr Asn 565 570 575 Gly Ser Leu Leu Leu Ile Val Gln Asp Gly Val Gly Gly Leu Tyr Gln 580 585 590 Cys Trp Ala Thr Glu Asn Gly Phe Ser Tyr Pro Val Ile Ser Tyr Trp 595 600 605 Val Asp Ser Gln Asp Gln Thr Leu Ala Leu Asp Pro Glu Leu Ala Gly 610 615 620 Ile Pro Arg Glu His Val Lys Val Pro Leu Thr Arg Val Ser Gly Gly 625 630 635 640 Ala Ala Leu Ala Ala Gln Gln Ser Tyr Trp Pro His 645 650 23 21 PRT Homo sapiens 23 Phe Val Thr Val Thr Val Leu Phe Ala Leu Val Leu Ser Gly Ala Leu 1 5 10 15 Ile Ile Leu Val Ala 20 24 57 PRT Homo sapiens 24 Ser Pro Leu Arg Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Glu 1 5 10 15 Thr Leu Arg Pro Gly Glu Lys Ala Pro Leu Ser Arg Glu Gln His Leu 20 25 30 Gln Ser Pro Lys Glu Cys Arg Thr Ser Ala Ser Asp Val Asp Ala Asp 35 40 45 Asn Asn Cys Leu Gly Thr Glu Val Ala 50 55 25 2964 DNA Homo sapiens 25 gtcgacccac gcgtccgcgg acgcgtgggg acggctcccg gctgcagtct gcccgcccgc 60 cccgcgcggg ggccgagtcg cgaagcgcgc ctgcgacccg gcgtccgggc gcgctggaga 120 ggacgcgagg agccatgagg cgccagcctg cgaaggtggc ggcgctgctg ctcgggctgc 180 tcttggagtg cacagaagcc aaaaagcatt gctggtattt cgaaggactc tatccaacct 240 attatatatg ccgctcctac gaggactgct gtggctccag gtgctgtgtg cgggccctct 300 ccatacagag gctgtggtac ttctggttcc ttctgatgat gggcgtgctt ttctgctgcg 360 gagccggctt cttcatccgg aggcgcatgt accccccgcc gctgatcgag gagccagcct 420 tcaatgtgtc ctacaccagg cagcccccaa atcccggccc aggagcccag cagccggggc 480 cgccctatta cactgaccca ggaggaccgg ggatgaaccc tgtcgggaat tccatggcaa 540 tggctttcca ggtcccaccc aactcacccc aggggagtgt ggcctgcccg ccccctccag 600 cctactgcaa cacgcctccg cccccgtacg aacaggtagt gaaggccaag tagtggggtg 660 cccacgtgca agaggagaga caggagaggg cctttccctg gcctttctgt cttcgttgat 720 gttcacttcc aggaacggtc tcgtgggctg ctaagggcag ttcctctgat atcctcacag 780 caagcacagc tctctttcag gctttccatg gagtacaata tatgaactca cactttgtct 840 cctctgttgc ttctgtttct gacgcagtct gtgctctcac atggtagtgt ggtgacagtc 900 cccgagggct gacgtcctta cggtggcgtg accagatcta caggagagag actgagagga 960 agaaggcagt gctggaggtg caggtggcat gtagaggggc caggccgagc atcccaggca 1020 agcatccttc tgcccgggta ttaataggaa gccccatgcc gggcggctca gccgatgaag 1080 cagcagccga ctgagctgag cccagcaggt catctgctcc agcctgtcct ctcgtcagcc 1140 ttcctcttcc agaagctgtt ggagagacat tcaggagaga gcaagcccct tgtcatgttt 1200 ctgtctctgt tcatatccta aagatagact tctcctgcac cgccagggaa gggtagcacg 1260 tgcagctctc accgcaggat ggggcctaga atcaggcttg ccttggaggc ctgacagtga 1320 tctgacatcc actaagcaaa tttatttaaa ttcatgggaa atcacttcct gccccaaact 1380 gagacattgc attttgtgag ctcttggtct gatttggaga aaggactgtt acccattttt 1440 ttggtgtgtt tatggaagtg catgtagagc gtcctgccct ttgaaatcag actgggtgtg 1500 tgtcttccct ggacatcact gcctctccag ggcattctca ggcccggggg tctccttccc 1560 tcaggcagct ccagtggtgg gttctgaagg gtgctttcaa aacggggcac atctggctgg 1620 gaagtcacat ggactcttcc agggagagag accagctgag gcgtctctct ctgaggttgt 1680 gttgggtcta agcgggtgtg tgctgggctc caaggaggag gagcttgctg ggaaaagaca 1740 ggagaagtac tgactcaact gcactgacca tgttgtcata attagaataa agaagaagtg 1800 gtcggaaatg cacattcctg gataggaatc acagctcacc ccaggatctc acaggtagtc 1860 tcctgagtag ttgacggcta gcggggagct agttccgccg catagttata gtgttgatgt 1920 gtgaacgctg acctgtcctg tgtgctaaga gctatgcagc ttagctgagg cgcctagatt 1980 actagatgtg ctgtatcacg gggaatgagg tgggggtgct tattttttaa tgaactaatc 2040 agagcctctt gagaaattgt tactcattga actggagcat caagacatct catggaagtg 2100 gatacggagt gatttggtgt ccatgctttt cactctgagg acatttaatc ggagaacctc 2160 ctggggaatt ttgtgggaga cacttgggaa caaaacagac accctgggaa tgcagttgca 2220 agcacagatg ctgccaccag tgtctctgac caccctggtg tgactgctga ctgccagcgt 2280 ggtacctccc atgctgcagg cctccatcta aatgagacaa caaagcacaa tgttcactgt 2340 ttacaaccaa gacaactgcg tgggtccaaa cactcctctt cctccaggtc atttgttttg 2400 catttttaat gtctttattt tttgtaatga aaaagcacac taagctgccc ctggaatcgg 2460 gtgcagctga ataggcaccc aaaagtccgt gactaaattt cgtttgtctt tttgatagca 2520 aattatgtta agagacagtg atggctaggg ctcaacaatt ttgtattccc atgtttgtgt 2580 gagacagagt ttgttttccc ttgaacttgg ttagaattgt gctactgtga acgctgatcc 2640 tgcatatgga agtcccactt tggtgacatt tcctggccat tcttgtttcc attgtgtgga 2700 tggtgggttg tgcccacttc ctggagtgag acagctcctg gtgtgtagaa ttcccggagc 2760 gtccgtggtt cagagtaaac ttgaagcaga tctgtgcatg cttttcctct gcaacaattg 2820 gctcgtttct cttttttgtt ctcttttgat aggatcctgt ttcctatgtg tgcaaaataa 2880 aaataaattt gggcaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2940 aaaaaaaaaa aaaagggcgg ccgc 2964 26 516 DNA Homo sapiens 26 atgaggcgcc agcctgcgaa ggtggcggcg ctgctgctcg ggctgctctt ggagtgcaca 60 gaagccaaaa agcattgctg gtatttcgaa ggactctatc caacctatta tatatgccgc 120 tcctacgagg actgctgtgg ctccaggtgc tgtgtgcggg ccctctccat acagaggctg 180 tggtacttct ggttccttct gatgatgggc gtgcttttct gctgcggagc cggcttcttc 240 atccggaggc gcatgtaccc cccgccgctg atcgaggagc cagccttcaa tgtgtcctac 300 accaggcagc ccccaaatcc cggcccagga gcccagcagc cggggccgcc ctattacact 360 gacccaggag gaccggggat gaaccctgtc gggaattcca tggcaatggc tttccaggtc 420 ccacccaact caccccaggg gagtgtggcc tgcccgcccc ctccagccta ctgcaacacg 480 cctccgcccc cgtacgaaca ggtagtgaag gccaag 516 27 172 PRT Homo sapiens 27 Met Arg Arg Gln Pro Ala Lys Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Leu Glu Cys Thr Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30 Tyr Pro Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45 Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp 50 55 60 Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly Ala Gly Phe Phe 65 70 75 80 Ile Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro Ala Phe 85 90 95 Asn Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln 100 105 110 Gln Pro Gly Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro Val Gly Asn Ser Met Ala Met Ala Phe Gln Val Pro Pro Asn Ser 130 135 140 Pro Gln Gly Ser Val Ala Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr 145 150 155 160 Pro Pro Pro Pro Tyr Glu Gln Val Val Lys Ala Lys 165 170 28 22 PRT Homo sapiens 28 Met Arg Arg Gln Pro Ala Lys Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Leu Glu Cys Thr Glu Ala 20 29 150 PRT Homo sapiens 29 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly 35 40 45 Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr 50 55 60 Pro Pro Pro Leu Ile Glu Glu Pro Ala Phe Asn Val Ser Tyr Thr Arg 65 70 75 80 Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln Gln Pro Gly Pro Pro Tyr 85 90 95 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro Val Gly Asn Ser Met 100 105 110 Ala Met Ala Phe Gln Val Pro Pro Asn Ser Pro Gln Gly Ser Val Ala 115 120 125 Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr Pro Pro Pro Pro Tyr Glu 130 135 140 Gln Val Val Lys Ala Lys 145 150 30 38 PRT Homo sapiens 30 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser Ile Gln Arg Leu 35 31 21 PRT Homo sapiens 31 Trp Tyr Phe Trp Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly 1 5 10 15 Ala Gly Phe Phe Ile 20 32 91 PRT Homo sapiens 32 Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro Ala Phe Asn 1 5 10 15 Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln Gln 20 25 30 Pro Gly Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro 35 40 45 Val Gly Asn Ser Met Ala Met Ala Phe Gln Val Pro Pro Asn Ser Pro 50 55 60 Gln Gly Ser Val Ala Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr Pro 65 70 75 80 Pro Pro Pro Tyr Glu Gln Val Val Lys Ala Lys 85 90 33 1980 DNA Homo sapiens 33 gtcgacccac gcgtccgcag ctttggacac ttcctctgct tgaggacacc ttgactaacc 60 tccaagggca actaaaggat caagaaaggc ccagcacagc agaagatcag ctggatctag 120 ctcctgcagg agatgtgtac aaagacaatc ccagtcctct ggggatgttt cctcctgtgg 180 aatctctatg tctcatcctc tcagaccatt taccctggaa tcaaggcaag gattactcag 240 agggcacttg actatggtgt tcaagctgga atgaagatga ttgagcaaat gctaaaagaa 300 aagaaactcc cagatttaag cggttctgag tctcttgaat ttctaaaagt tgattatgta 360 aactacaatt tttcaaatat aaaaatcagt gccttttcat ttccaaatac ctcattggct 420 tttgtgcctg gagtgggaat caaagcgcta accaaccatg gcactgccaa catcagcaca 480 gactgggggt tcgagtctcc actttttgtt ctgtataact cctttgctga gcccatggag 540 aaacccattt taaagaactt aaatgaaatg ctctgtccca ttattgcaag tgaagtcaaa 600 gcgctaaatg ccaacctcag cacactggag gttttaacca agattgacaa ctacactctg 660 ctggattact ccctaatcag ttctccagaa attactgaga actaccttga cctgaacttg 720 aagggtgtat tctacccact ggaaaacctc accgaccccc ccttctcacc agttcctttt 780 gtgctcccag aacgcagcaa ctccatgctc tacattggaa tcgccgagta tttctttaaa 840 tctgcgtcct ttgctcattt cacagctggg gttttcaatc tcactctctc caccgaagag 900 atttccaacc attttgttca aaactctcaa ggccttggca acgtgctctc ccggattgca 960 gagatctaca tcttgtccca gcccttcatg gtgaggatca tggccacaga gcctcccata 1020 atcaatctac aaccaggcaa tttcaccctg gacatccctg cctccatcat gatgctcacc 1080 caacccaaga actccacagt tgaaaccatc gtttccatgg acttcgttgc tagtaccagt 1140 gttggcctgg ttattttggg acaaagactg gtctgctcct tgtctctgaa cagattccgc 1200 cttgctttgc cagagtccaa tcgcagcaac attgaggtct tgaggtttga aaatattcta 1260 tcgtccattc ttcactttgg agtcctccca ctggccaatg caaaattgca gcaaggattt 1320 cctctgccca atccacacaa attcttattc gtcaattcag atattgaagt tcttgagggt 1380 ttccttttga tttccaccga cctgaagtat gaaacatcct caaagcagca gccaagtttc 1440 cacgtatggg aaggtctgaa cctgataagc agacagtgga gggggaagtc agccccttga 1500 ttgccggttt gcaattcacc ccaggaagta aatggtcctt aatcctacaa ctactgtaaa 1560 cccagaaggg aaagacagta cacactggaa ttgtaaagcc cttgtgaatt gcttaggcag 1620 aaagttttct ttcttaagcc ttcaggaacc cagaataagg cagactctgt taaagggata 1680 aatagaggtg tctgaatgtg agtgtatgca tgctgcgtgt gtctgtgttt atgtttgttt 1740 gtttgtttgg ggcaagaaag attctaggac aagagctagg catgtacttc tgaccaggtg 1800 ggtaagcaac tctaagtctg tatttgtatt ggtcattctc agtggaaatc ccttaggccc 1860 tctagtggtt ttcccctacc tgcatattgg ttttcatgtt ttatattcac tgttactatc 1920 ttctgtgttt aattaaaatt gttttctatc aaaaaaaaaa aaaaaaaaaa gggcggccgc 1980 34 1365 DNA Homo sapiens 34 atgtgtacaa agacaatccc agtcctctgg ggatgtttcc tcctgtggaa tctctatgtc 60 tcatcctctc agaccattta ccctggaatc aaggcaagga ttactcagag ggcacttgac 120 tatggtgttc aagctggaat gaagatgatt gagcaaatgc taaaagaaaa gaaactccca 180 gatttaagcg gttctgagtc tcttgaattt ctaaaagttg attatgtaaa ctacaatttt 240 tcaaatataa aaatcagtgc cttttcattt ccaaatacct cattggcttt tgtgcctgga 300 gtgggaatca aagcgctaac caaccatggc actgccaaca tcagcacaga ctgggggttc 360 gagtctccac tttttgttct gtataactcc tttgctgagc ccatggagaa acccatttta 420 aagaacttaa atgaaatgct ctgtcccatt attgcaagtg aagtcaaagc gctaaatgcc 480 aacctcagca cactggaggt tttaaccaag attgacaact acactctgct ggattactcc 540 ctaatcagtt ctccagaaat tactgagaac taccttgacc tgaacttgaa gggtgtattc 600 tacccactgg aaaacctcac cgaccccccc ttctcaccag ttccttttgt gctcccagaa 660 cgcagcaact ccatgctcta cattggaatc gccgagtatt tctttaaatc tgcgtccttt 720 gctcatttca cagctggggt tttcaatctc actctctcca ccgaagagat ttccaaccat 780 tttgttcaaa actctcaagg ccttggcaac gtgctctccc ggattgcaga gatctacatc 840 ttgtcccagc ccttcatggt gaggatcatg gccacagagc ctcccataat caatctacaa 900 ccaggcaatt tcaccctgga catccctgcc tccatcatga tgctcaccca acccaagaac 960 tccacagttg aaaccatcgt ttccatggac ttcgttgcta gtaccagtgt tggcctggtt 1020 attttgggac aaagactggt ctgctccttg tctctgaaca gattccgcct tgctttgcca 1080 gagtccaatc gcagcaacat tgaggtcttg aggtttgaaa atattctatc gtccattctt 1140 cactttggag tcctcccact ggccaatgca aaattgcagc aaggatttcc tctgcccaat 1200 ccacacaaat tcttattcgt caattcagat attgaagttc ttgagggttt ccttttgatt 1260 tccaccgacc tgaagtatga aacatcctca aagcagcagc caagtttcca cgtatgggaa 1320 ggtctgaacc tgataagcag acagtggagg gggaagtcag cccct 1365 35 455 PRT Homo sapiens 35 Met Cys Thr Lys Thr Ile Pro Val Leu Trp Gly Cys Phe Leu Leu Trp 1 5 10 15 Asn Leu Tyr Val Ser Ser Ser Gln Thr Ile Tyr Pro Gly Ile Lys Ala 20 25 30 Arg Ile Thr Gln Arg Ala Leu Asp Tyr Gly Val Gln Ala Gly Met Lys 35 40 45 Met Ile Glu Gln Met Leu Lys Glu Lys Lys Leu Pro Asp Leu Ser Gly 50 55 60 Ser Glu Ser Leu Glu Phe Leu Lys Val Asp Tyr Val Asn Tyr Asn Phe 65 70 75 80 Ser Asn Ile Lys Ile Ser Ala Phe Ser Phe Pro Asn Thr Ser Leu Ala 85 90 95 Phe Val Pro Gly Val Gly Ile Lys Ala Leu Thr Asn His Gly Thr Ala 100 105 110 Asn Ile Ser Thr Asp Trp Gly Phe Glu Ser Pro Leu Phe Val Leu Tyr 115 120 125 Asn Ser Phe Ala Glu Pro Met Glu Lys Pro Ile Leu Lys Asn Leu Asn 130 135 140 Glu Met Leu Cys Pro Ile Ile Ala Ser Glu Val Lys Ala Leu Asn Ala 145 150 155 160 Asn Leu Ser Thr Leu Glu Val Leu Thr Lys Ile Asp Asn Tyr Thr Leu 165 170 175 Leu Asp Tyr Ser Leu Ile Ser Ser Pro Glu Ile Thr Glu Asn Tyr Leu 180 185 190 Asp Leu Asn Leu Lys Gly Val Phe Tyr Pro Leu Glu Asn Leu Thr Asp 195 200 205 Pro Pro Phe Ser Pro Val Pro Phe Val Leu Pro Glu Arg Ser Asn Ser 210 215 220 Met Leu Tyr Ile Gly Ile Ala Glu Tyr Phe Phe Lys Ser Ala Ser Phe 225 230 235 240 Ala His Phe Thr Ala Gly Val Phe Asn Leu Thr Leu Ser Thr Glu Glu 245 250 255 Ile Ser Asn His Phe Val Gln Asn Ser Gln Gly Leu Gly Asn Val Leu 260 265 270 Ser Arg Ile Ala Glu Ile Tyr Ile Leu Ser Gln Pro Phe Met Val Arg 275 280 285 Ile Met Ala Thr Glu Pro Pro Ile Ile Asn Leu Gln Pro Gly Asn Phe 290 295 300 Thr Leu Asp Ile Pro Ala Ser Ile Met Met Leu Thr Gln Pro Lys Asn 305 310 315 320 Ser Thr Val Glu Thr Ile Val Ser Met Asp Phe Val Ala Ser Thr Ser 325 330 335 Val Gly Leu Val Ile Leu Gly Gln Arg Leu Val Cys Ser Leu Ser Leu 340 345 350 Asn Arg Phe Arg Leu Ala Leu Pro Glu Ser Asn Arg Ser Asn Ile Glu 355 360 365 Val Leu Arg Phe Glu Asn Ile Leu Ser Ser Ile Leu His Phe Gly Val 370 375 380 Leu Pro Leu Ala Asn Ala Lys Leu Gln Gln Gly Phe Pro Leu Pro Asn 385 390 395 400 Pro His Lys Phe Leu Phe Val Asn Ser Asp Ile Glu Val Leu Glu Gly 405 410 415 Phe Leu Leu Ile Ser Thr Asp Leu Lys Tyr Glu Thr Ser Ser Lys Gln 420 425 430 Gln Pro Ser Phe His Val Trp Glu Gly Leu Asn Leu Ile Ser Arg Gln 435 440 445 Trp Arg Gly Lys Ser Ala Pro 450 455 36 23 PRT Homo sapiens 36 Met Cys Thr Lys Thr Ile Pro Val Leu Trp Gly Cys Phe Leu Leu Trp 1 5 10 15 Asn Leu Tyr Val Ser Ser Ser 20 37 432 PRT Homo sapiens 37 Gln Thr Ile Tyr Pro Gly Ile Lys Ala Arg Ile Thr Gln Arg Ala Leu 1 5 10 15 Asp Tyr Gly Val Gln Ala Gly Met Lys Met Ile Glu Gln Met Leu Lys 20 25 30 Glu Lys Lys Leu Pro Asp Leu Ser Gly Ser Glu Ser Leu Glu Phe Leu 35 40 45 Lys Val Asp Tyr Val Asn Tyr Asn Phe Ser Asn Ile Lys Ile Ser Ala 50 55 60 Phe Ser Phe Pro Asn Thr Ser Leu Ala Phe Val Pro Gly Val Gly Ile 65 70 75 80 Lys Ala Leu Thr Asn His Gly Thr Ala Asn Ile Ser Thr Asp Trp Gly 85 90 95 Phe Glu Ser Pro Leu Phe Val Leu Tyr Asn Ser Phe Ala Glu Pro Met 100 105 110 Glu Lys Pro Ile Leu Lys Asn Leu Asn Glu Met Leu Cys Pro Ile Ile 115 120 125 Ala Ser Glu Val Lys Ala Leu Asn Ala Asn Leu Ser Thr Leu Glu Val 130 135 140 Leu Thr Lys Ile Asp Asn Tyr Thr Leu Leu Asp Tyr Ser Leu Ile Ser 145 150 155 160 Ser Pro Glu Ile Thr Glu Asn Tyr Leu Asp Leu Asn Leu Lys Gly Val 165 170 175 Phe Tyr Pro Leu Glu Asn Leu Thr Asp Pro Pro Phe Ser Pro Val Pro 180 185 190 Phe Val Leu Pro Glu Arg Ser Asn Ser Met Leu Tyr Ile Gly Ile Ala 195 200 205 Glu Tyr Phe Phe Lys Ser Ala Ser Phe Ala His Phe Thr Ala Gly Val 210 215 220 Phe Asn Leu Thr Leu Ser Thr Glu Glu Ile Ser Asn His Phe Val Gln 225 230 235 240 Asn Ser Gln Gly Leu Gly Asn Val Leu Ser Arg Ile Ala Glu Ile Tyr 245 250 255 Ile Leu Ser Gln Pro Phe Met Val Arg Ile Met Ala Thr Glu Pro Pro 260 265 270 Ile Ile Asn Leu Gln Pro Gly Asn Phe Thr Leu Asp Ile Pro Ala Ser 275 280 285 Ile Met Met Leu Thr Gln Pro Lys Asn Ser Thr Val Glu Thr Ile Val 290 295 300 Ser Met Asp Phe Val Ala Ser Thr Ser Val Gly Leu Val Ile Leu Gly 305 310 315 320 Gln Arg Leu Val Cys Ser Leu Ser Leu Asn Arg Phe Arg Leu Ala Leu 325 330 335 Pro Glu Ser Asn Arg Ser Asn Ile Glu Val Leu Arg Phe Glu Asn Ile 340 345 350 Leu Ser Ser Ile Leu His Phe Gly Val Leu Pro Leu Ala Asn Ala Lys 355 360 365 Leu Gln Gln Gly Phe Pro Leu Pro Asn Pro His Lys Phe Leu Phe Val 370 375 380 Asn Ser Asp Ile Glu Val Leu Glu Gly Phe Leu Leu Ile Ser Thr Asp 385 390 395 400 Leu Lys Tyr Glu Thr Ser Ser Lys Gln Gln Pro Ser Phe His Val Trp 405 410 415 Glu Gly Leu Asn Leu Ile Ser Arg Gln Trp Arg Gly Lys Ser Ala Pro 420 425 430 38 483 PRT Homo sapiens 38 Met Ala Arg Gly Pro Cys Asn Ala Pro Arg Trp Val Ser Leu Met Val 1 5 10 15 Leu Val Ala Ile Gly Thr Ala Val Thr Ala Ala Val Asn Pro Gly Val 20 25 30 Val Val Arg Ile Ser Gln Lys Gly Leu Asp Tyr Ala Ser Gln Gln Gly 35 40 45 Thr Ala Ala Leu Gln Lys Glu Leu Lys Arg Ile Lys Ile Pro Asp Tyr 50 55 60 Ser Asp Ser Phe Lys Ile Lys His Leu Gly Lys Gly His Tyr Ser Phe 65 70 75 80 Tyr Ser Met Asp Ile Arg Glu Phe Gln Leu Pro Ser Ser Gln Ile Ser 85 90 95 Met Val Pro Asn Val Gly Leu Lys Phe Ser Ile Ser Asn Ala Asn Ile 100 105 110 Lys Ile Ser Gly Lys Trp Lys Ala Gln Lys Arg Phe Leu Lys Met Ser 115 120 125 Gly Asn Phe Asp Leu Ser Ile Glu Gly Met Ser Ile Ser Ala Asp Leu 130 135 140 Lys Leu Gly Ser Asn Pro Thr Ser Gly Lys Pro Thr Ile Thr Cys Ser 145 150 155 160 Ser Cys Ser Ser His Ile Asn Ser Val His Val His Ile Ser Lys Ser 165 170 175 Lys Val Gly Trp Leu Ile Gln Leu Phe His Lys Lys Ile Glu Ser Ala 180 185 190 Leu Arg Asn Lys Met Asn Ser Gln Val Cys Glu Lys Val Thr Asn Ser 195 200 205 Val Ser Ser Lys Leu Gln Pro Tyr Phe Gln Thr Leu Pro Val Met Thr 210 215 220 Lys Ile Asp Ser Val Ala Gly Ile Asn Tyr Gly Leu Val Ala Pro Pro 225 230 235 240 Ala Thr Thr Ala Glu Thr Leu Asp Val Gln Met Lys Gly Glu Phe Tyr 245 250 255 Ser Glu Asn His His Asn Pro Pro Pro Phe Ala Pro Pro Val Met Glu 260 265 270 Phe Pro Ala Ala His Asp Arg Met Val Tyr Leu Gly Leu Ser Asp Tyr 275 280 285 Phe Phe Asn Thr Ala Gly Leu Val Tyr Gln Glu Ala Gly Val Leu Lys 290 295 300 Met Thr Leu Arg Asp Asp Met Ile Pro Lys Glu Ser Lys Phe Arg Leu 305 310 315 320 Thr Thr Lys Phe Phe Gly Thr Phe Leu Pro Glu Val Ala Lys Lys Phe 325 330 335 Pro Asn Met Lys Ile Gln Ile His Val Ser Ala Ser Thr Pro Pro His 340 345 350 Leu Ser Val Gln Pro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val 355 360 365 Gln Ala Phe Ala Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu 370 375 380 Ile Gly Met His Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn 385 390 395 400 Arg Leu Val Gly Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu Leu Lys 405 410 415 His Ser Asn Ile Gly Pro Phe Pro Val Glu Leu Leu Gln Asp Ile Met 420 425 430 Asn Tyr Ile Val Pro Ile Leu Val Leu Pro Arg Val Asn Glu Lys Leu 435 440 445 Gln Lys Gly Phe Pro Leu Pro Thr Pro Ala Arg Val Gln Leu Tyr Asn 450 455 460 Val Val Leu Gln Pro His Gln Asn Phe Leu Leu Phe Gly Ala Asp Val 465 470 475 480 Val Tyr Lys 39 481 PRT Homo sapiens 39 Met Gly Ala Leu Ala Arg Ala Leu Pro Ser Ile Leu Leu Ala Leu Leu 1 5 10 15 Leu Thr Ser Thr Pro Glu Ala Leu Gly Ala Asn Pro Gly Leu Val Ala 20 25 30 Arg Ile Thr Asp Lys Gly Leu Gln Tyr Ala Ala Gln Glu Gly Leu Leu 35 40 45 Ala Leu Gln Ser Glu Leu Leu Arg Ile Thr Leu Pro Asp Phe Thr Gly 50 55 60 Asp Leu Arg Ile Pro His Val Gly Arg Gly Arg Tyr Glu Phe His Ser 65 70 75 80 Leu Asn Ile His Glu Phe Gln Leu Pro Ser Ser Gln Ile Ser Met Val 85 90 95 Pro Asn Val Gly Leu Lys Phe Ser Ile Ser Asn Ala Asn Ile Lys Ile 100 105 110 Ser Gly Lys Trp Lys Ala Gln Lys Arg Phe Leu Lys Met Ser Gly Asn 115 120 125 Phe Asp Leu Ser Ile Glu Gly Met Ser Ile Ser Ala Asp Leu Lys Leu 130 135 140 Gly Ser Asn Pro Thr Ser Gly Lys Pro Thr Ile Thr Cys Ser Ser Cys 145 150 155 160 Ser Ser His Ile Asn Ser Val His Val His Ile Ser Lys Ser Lys Val 165 170 175 Gly Trp Leu Ile Gln Leu Phe His Lys Lys Ile Glu Ser Ala Leu Arg 180 185 190 Asn Lys Met Asn Ser Gln Val Cys Glu Lys Val Thr Asn Ser Val Ser 195 200 205 Ser Lys Leu Gln Pro Tyr Phe Gln Thr Leu Pro Val Met Thr Lys Ile 210 215 220 Asp Ser Val Ala Gly Ile Asn Tyr Gly Leu Val Ala Pro Pro Ala Thr 225 230 235 240 Thr Ala Glu Thr Leu Asp Val Gln Met Lys Gly Glu Phe Tyr Ser Glu 245 250 255 Asn His His Asn Pro Pro Pro Phe Ala Pro Pro Val Met Glu Phe Pro 260 265 270 Ala Ala His Asp Arg Met Val Tyr Leu Gly Leu Ser Asp Tyr Phe Phe 275 280 285 Asn Thr Ala Gly Leu Val Tyr Gln Glu Ala Gly Val Leu Lys Met Thr 290 295 300 Leu Arg Asp Asp Met Ile Pro Lys Glu Ser Lys Phe Arg Leu Thr Thr 305 310 315 320 Lys Phe Phe Gly Thr Phe Leu Pro Glu Val Ala Lys Lys Phe Pro Asn 325 330 335 Met Lys Ile Gln Ile His Val Ser Ala Ser Thr Pro Pro His Leu Ser 340 345 350 Val Gln Pro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val Gln Ala 355 360 365 Leu Ala Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu Ile Gly 370 375 380 Met His Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn Arg Leu 385 390 395 400 Val Gly Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu Leu Lys His Ser 405 410 415 Asn Ile Gly Pro Phe Pro Val Glu Leu Leu Gln Asp Ile Met Asn Tyr 420 425 430 Ile Val Pro Ile Leu Val Leu Pro Arg Val Asn Glu Lys Leu Gln Lys 435 440 445 Gly Phe Pro Leu Pro Thr Pro Ala Arg Val Gln Leu Tyr Asn Val Val 450 455 460 Leu Gln Pro His Gln Asn Phe Leu Leu Phe Gly Ala Asp Val Val Tyr 465 470 475 480 Lys 40 383 PRT Caenorhabditis elegans 40 Met Arg Ile Ala His Ala Ser Ser Arg Gly Asn Ile Ser Ile Phe Ser 1 5 10 15 Val Phe Leu Ile Pro Leu Ile Ala Tyr Ile Leu Ile Leu Pro Gly Val 20 25 30 Arg Arg Lys Arg Val Val Thr Thr Val Thr Tyr Val Leu Met Leu Ala 35 40 45 Val Gly Gly Ala Leu Ile Ala Ser Leu Ile Tyr Pro Cys Trp Ala Ser 50 55 60 Gly Ser Gln Met Ile Tyr Thr Gln Phe Arg Gly His Ser Asn Glu Arg 65 70 75 80 Ile Leu Ala Lys Ile Gly Val Glu Ile Gly Leu Gln Lys Val Asn Val 85 90 95 Thr Leu Lys Phe Glu Arg Leu Leu Ser Ser Asn Asp Val Leu Pro Gly 100 105 110 Ser Asp Met Thr Glu Leu Tyr Tyr Asn Glu Gly Phe Asp Ile Ser Gly 115 120 125 Ile Ser Ser Met Ala Glu Ala Leu His His Gly Leu Glu Asn Gly Leu 130 135 140 Pro Tyr Pro Met Leu Ser Val Leu Glu Tyr Phe Ser Leu Asn Gln Asp 145 150 155 160 Ser Phe Asp Trp Gly Arg His Tyr Arg Val Ala Gly His Tyr Thr His 165 170 175 Ala Ala Ile Trp Phe Ala Phe Ala Cys Trp Cys Leu Ser Val Val Leu 180 185 190 Met Leu Phe Leu Pro His Asn Ala Tyr Lys Ser Ile Leu Ala Thr Gly 195 200 205 Ile Ser Cys Leu Ile Ala Cys Leu Val Tyr Leu Leu Leu Ser Pro Cys 210 215 220 Glu Leu Arg Ile Ala Phe Thr Gly Glu Asn Phe Glu Arg Val Asp Leu 225 230 235 240 Thr Ala Thr Phe Ser Phe Cys Phe Tyr Leu Ile Phe Ala Ile Gly Ile 245 250 255 Leu Cys Val Leu Cys Gly Leu Gly Leu Gly Ile Cys Glu His Trp Arg 260 265 270 Ile Tyr Thr Leu Ser Thr Phe Leu Asp Ala Ser Leu Asp Glu His Val 275 280 285 Gly Pro Lys Trp Lys Lys Leu Pro Thr Gly Gly Pro Ala Leu Gln Gly 290 295 300 Val Gln Ile Gly Ala Tyr Gly Thr Asn Thr Thr Asn Ser Ser Arg Asp 305 310 315 320 Lys Asn Asp Ile Ser Ser Asp Lys Thr Ala Gly Ser Ser Gly Phe Gln 325 330 335 Ser Arg Thr Ser Thr Cys Gln Ser Ser Ala Ser Ser Ala Ser Leu Arg 340 345 350 Ser Gln Ser Ser Ile Glu Thr Val His Asp Glu Ala Glu Leu Glu Arg 355 360 365 Thr His Val His Phe Leu Gln Glu Pro Cys Ser Ser Ser Ser Thr 370 375 380 41 399 PRT Homo sapiens 41 Met Lys Met Arg Phe Leu Gly Leu Val Val Cys Leu Val Leu Trp Pro 1 5 10 15 Leu His Ser Glu Gly Ser Gly Gly Lys Leu Thr Ala Val Asp Pro Glu 20 25 30 Thr Asn Met Asn Val Ser Glu Ile Ile Ser Tyr Trp Gly Phe Pro Ser 35 40 45 Glu Glu Tyr Leu Val Glu Thr Glu Asp Gly Tyr Ile Leu Cys Leu Asn 50 55 60 Arg Ile Pro His Gly Arg Lys Asn His Ser Asp Lys Gly Pro Lys Pro 65 70 75 80 Val Val Phe Leu Gln His Gly Leu Leu Ala Asp Ser Ser Asn Trp Val 85 90 95 Thr Asn Leu Ala Asn Ser Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly 100 105 110 Phe Asp Val Trp Met Gly Asn Ser Arg Gly Asn Thr Trp Ser Arg Lys 115 120 125 His Lys Thr Leu Ser Val Ser Gln Asp Glu Phe Trp Ala Phe Ser Tyr 130 135 140 Asp Glu Met Ala Lys Tyr Asp Leu Pro Ala Ser Ile Asn Phe Ile Leu 145 150 155 160 Asn Lys Thr Gly Gln Glu Gln Val Tyr Tyr Val Gly His Ser Gln Gly 165 170 175 Thr Thr Ile Gly Phe Ile Ala Phe Ser Gln Ile Pro Glu Leu Ala Lys 180 185 190 Arg Ile Lys Met Phe Phe Ala Leu Gly Pro Val Ala Ser Val Ala Phe 195 200 205 Cys Thr Ser Pro Met Ala Lys Leu Gly Arg Leu Pro Asp His Leu Ile 210 215 220 Lys Asp Leu Phe Gly Asp Lys Glu Phe Leu Pro Gln Ser Ala Phe Leu 225 230 235 240 Lys Trp Leu Gly Thr His Val Cys Thr His Val Ile Leu Lys Glu Leu 245 250 255 Cys Gly Asn Leu Cys Phe Leu Leu Cys Gly Phe Asn Glu Arg Asn Leu 260 265 270 Asn Met Ser Arg Val Asp Val Tyr Thr Thr His Ser Pro Ala Gly Thr 275 280 285 Ser Val Gln Asn Met Leu His Trp Ser Gln Ala Val Lys Phe Gln Lys 290 295 300 Phe Gln Ala Phe Asp Trp Gly Ser Ser Ala Lys Asn Tyr Phe His Tyr 305 310 315 320 Asn Gln Ser Tyr Pro Pro Thr Tyr Asn Val Lys Asp Met Leu Val Pro 325 330 335 Thr Ala Val Trp Ser Gly Gly His Asp Trp Leu Ala Asp Val Tyr Asp 340 345 350 Val Asn Ile Leu Leu Thr Gln Ile Thr Asn Leu Val Phe His Glu Ser 355 360 365 Ile Pro Glu Trp Glu His Leu Asp Phe Ile Trp Gly Leu Asp Ala Pro 370 375 380 Trp Arg Leu Tyr Asn Lys Ile Ile Asn Leu Met Arg Lys Tyr Gln 385 390 395 42 19 PRT Mus sp. 42 Met Ala Pro Pro Ala Ala Arg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala 43 451 PRT Mus sp. 43 Ala Arg Pro Ala Pro Gly Pro Arg Ser Gly Pro Glu Cys Phe Thr Ala 1 5 10 15 Asn Gly Ala Asp Tyr Arg Gly Thr Gln Ser Trp Thr Ala Leu Gln Gly 20 25 30 Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His Pro Tyr Asn 35 40 45 Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu His Asn Tyr 50 55 60 Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr Val Ala Glu 65 70 75 80 His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro Ala Cys Gln 85 90 95 Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn Pro Pro Pro 100 105 110 Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile Gln Thr Cys 115 120 125 Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly Met Glu Ser 130 135 140 Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp Tyr Trp Lys His Gly 145 150 155 160 Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys Phe Gly Asp His Thr 165 170 175 Gln Pro Cys Gly Gly Asp Gly Arg Ile Ile Leu Phe Asp Thr Leu Val 180 185 190 Gly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ala Ala Val Val Tyr Ser 195 200 205 Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg Val Cys Tyr Trp Thr 210 215 220 Ile Arg Val Pro Gly Ala Ser Arg Ile His Phe Asn Phe Thr Leu Phe 225 230 235 240 Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu Leu Asp Gly Tyr Thr 245 250 255 His Arg Val Leu Val Arg Leu Ser Gly Arg Ser Arg Pro Pro Leu Ser 260 265 270 Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe Ser Asp Arg 275 280 285 Ile Asn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala Thr Lys Glu 290 295 300 Glu Pro Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Leu Ala Glu Val 305 310 315 320 Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala His Ser Ser Lys 325 330 335 Val Leu Tyr Val Ile Thr Pro Ser Pro Ser His Pro Pro Gln Thr Ala 340 345 350 Gln Val Ala Ile Pro Gly His Arg Gln Leu Gly Pro Thr Ala Thr Glu 355 360 365 Trp Lys Asp Gly Leu Cys Thr Ala Trp Arg Pro Ser Ser Ser Ser Gln 370 375 380 Ser Gln Gln Leu Ser Gln Arg Phe Phe Cys Met Ser His Leu Asn Leu 385 390 395 400 Ile Glu Ser Leu His Gln Glu Thr Leu Gly Thr Val Val Ser Leu Gly 405 410 415 Leu Leu Glu Ile Ser Gly Pro Phe Ser Met Asn Leu Pro Leu Gln Ser 420 425 430 Pro Ser Leu Arg Arg Ser Ser Arg Val Arg Val Asn Lys Met Thr Ala 435 440 445 Ile Pro Ser 450 44 150 PRT Mus sp. 44 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30 Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly 35 40 45 Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr 50 55 60 Pro Pro Pro Leu Ile Glu Glu Pro Thr Phe Asn Val Ser Tyr Thr Arg 65 70 75 80 Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln Gln Met Gly Pro Pro Tyr 85 90 95 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro Val Gly Asn Thr Met 100 105 110 Ala Met Ala Phe Gln Val Gln Pro Asn Ser Pro His Gly Gly Thr Thr 115 120 125 Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr Pro Pro Pro Pro Tyr Glu 130 135 140 Gln Val Val Lys Asp Lys 145 150 45 2044 DNA Homo sapiens 45 gtcgacccac gcgtccgggg aattgcagca ggaaaatatg tgaagagttt ttaaacccac 60 aaattcttct tactttagaa ttagttgtta cattggcagg aaaaaataaa tgcagatgtt 120 ggaccatgtt ggaaaccttg tcaagacagt ggattgtctc acacagaatg gaaatgtggc 180 ttctgattct ggtggcgtat atgttccaga gaaatgtgaa ttcagtacat atgccaacta 240 aagctgtgga cccagaagca ttcatgaata ttagtgaaat catccaacat caaggctatc 300 cctgtgagga atatgaagtc gcaactgaag atgggtatat cctttctgtt aacaggattc 360 ctcgaggcct agtgcaacct aagaagacag gttccaggcc tgtggtgtta ctgcagcatg 420 gcctagttgg aggtgctagc aactggattt ccaacctgcc caacaatagc ctgggcttca 480 ttctggcaga tgctggtttt gacgtgtgga tggggaacag caggggaaac gcctggtctc 540 gaaaacacaa gacactctcc atagaccaag atgagttctg ggctttcagt tatgatgaga 600 tggctaggtt tgaccttcct gcagtgataa actttatttt gcagaaaacg ggccaggaaa 660 agatctatta tgtcggctat tcacagggca ccaccatggg ctttattgca ttttccacca 720 tgccagagct ggctcagaaa atcaaaatgt attttgcttt agcacccata gccactgtta 780 agcatgcaaa aagccccggg accaaatttt tgttgctgcc agatatgatg atcaagggat 840 tgtttggcaa aaaagaattt ctgtatcaga ccagatttct cagacaactt gttatttacc 900 tttgtggcca ggtgattctt gatcagattt gtagtaatat catgttactt ctgggtggat 960 tcaacaccaa caatatgaac atgagccgag caagtgtata tgctgcccac actcttgctg 1020 gaacatctgt gcaaaatatt ctacactgga gccaggcagt gaattctggt gaactccggg 1080 catttgactg ggggagtgag accaaaaatc tggaaaaatg caatcagcca actcctgtaa 1140 ggtacagagt cagagatatg acggtcccta cagcaatgtg gacaggaggt caggactggc 1200 tttcaaatcc agaagacgtg aaaatgctgc tctctgaggt gaccaacctc atctaccata 1260 agaatattcc tgaatgggct cacgtggatt tcatctgggg tttggatgct cctcaccgta 1320 tgtacaatga aatcatccat ctgatgcagc aggaggagac caacctttcc cagggacggt 1380 gtgaggccgt attgtgaagc atctgacact gacgatctta ggacaacctc ctgagggatg 1440 gggctaggac ccatgaaggc agaattacgg agagcagaga cctagtatac atttttcaga 1500 ttccctgcac ttggcactaa atccgacact tacatttaca ttttttttct gtaaattaaa 1560 gtacttatta ggtaaataga ggttttgtat gctattatat attctaccat cttgaagggt 1620 aggttttacc tgatagccag aaaatatcta gacattctct atatcattca ggtaaatctc 1680 tttaaaacac ctattgtttt ttctataagc catatttttg gagcactaaa gtaaaatggc 1740 aaattgggac agatattgag gtctggagtc tgtggattat tgttgacttt gacaaaataa 1800 gctagacatt ttcaccttgt tgccacagag acataacact acctcaggaa gctgagctgc 1860 tttaaggaca acaacaacaa aatcagtgtt acagtatgga tgaaatctat gttaagcatt 1920 ctcagaataa ggccaagttt tatagttgca tctcagggaa gaaaatttta taggatgttt 1980 atgagttctc caataaatgc attctgcatt acataaaaaa aaaaaaaaaa aaaagggcgg 2040 ccgc 2044 46 1269 DNA Homo sapiens 46 atgttggaaa ccttgtcaag acagtggatt gtctcacaca gaatggaaat gtggcttctg 60 attctggtgg cgtatatgtt ccagagaaat gtgaattcag tacatatgcc aactaaagct 120 gtggacccag aagcattcat gaatattagt gaaatcatcc aacatcaagg ctatccctgt 180 gaggaatatg aagtcgcaac tgaagatggg tatatccttt ctgttaacag gattcctcga 240 ggcctagtgc aacctaagaa gacaggttcc aggcctgtgg tgttactgca gcatggccta 300 gttggaggtg ctagcaactg gatttccaac ctgcccaaca atagcctggg cttcattctg 360 gcagatgctg gttttgacgt gtggatgggg aacagcaggg gaaacgcctg gtctcgaaaa 420 cacaagacac tctccataga ccaagatgag ttctgggctt tcagttatga tgagatggct 480 aggtttgacc ttcctgcagt gataaacttt attttgcaga aaacgggcca ggaaaagatc 540 tattatgtcg gctattcaca gggcaccacc atgggcttta ttgcattttc caccatgcca 600 gagctggctc agaaaatcaa aatgtatttt gctttagcac ccatagccac tgttaagcat 660 gcaaaaagcc ccgggaccaa atttttgttg ctgccagata tgatgatcaa gggattgttt 720 ggcaaaaaag aatttctgta tcagaccaga tttctcagac aacttgttat ttacctttgt 780 ggccaggtga ttcttgatca gatttgtagt aatatcatgt tacttctggg tggattcaac 840 accaacaata tgaacatgag ccgagcaagt gtatatgctg cccacactct tgctggaaca 900 tctgtgcaaa atattctaca ctggagccag gcagtgaatt ctggtgaact ccgggcattt 960 gactggggga gtgagaccaa aaatctggaa aaatgcaatc agccaactcc tgtaaggtac 1020 agagtcagag atatgacggt ccctacagca atgtggacag gaggtcagga ctggctttca 1080 aatccagaag acgtgaaaat gctgctctct gaggtgacca acctcatcta ccataagaat 1140 attcctgaat gggctcacgt ggatttcatc tggggtttgg atgctcctca ccgtatgtac 1200 aatgaaatca tccatctgat gcagcaggag gagaccaacc tttcccaggg acggtgtgag 1260 gccgtattg 1269 47 423 PRT Homo sapiens 47 Met Leu Glu Thr Leu Ser Arg Gln Trp Ile Val Ser His Arg Met Glu 1 5 10 15 Met Trp Leu Leu Ile Leu Val Ala Tyr Met Phe Gln Arg Asn Val Asn 20 25 30 Ser Val His Met Pro Thr Lys Ala Val Asp Pro Glu Ala Phe Met Asn 35 40 45 Ile Ser Glu Ile Ile Gln His Gln Gly Tyr Pro Cys Glu Glu Tyr Glu 50 55 60 Val Ala Thr Glu Asp Gly Tyr Ile Leu Ser Val Asn Arg Ile Pro Arg 65 70 75 80 Gly Leu Val Gln Pro Lys Lys Thr Gly Ser Arg Pro Val Val Leu Leu 85 90 95 Gln His Gly Leu Val Gly Gly Ala Ser Asn Trp Ile Ser Asn Leu Pro 100 105 110 Asn Asn Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly Phe Asp Val Trp 115 120 125 Met Gly Asn Ser Arg Gly Asn Ala Trp Ser Arg Lys His Lys Thr Leu 130 135 140 Ser Ile Asp Gln Asp Glu Phe Trp Ala Phe Ser Tyr Asp Glu Met Ala 145 150 155 160 Arg Phe Asp Leu Pro Ala Val Ile Asn Phe Ile Leu Gln Lys Thr Gly 165 170 175 Gln Glu Lys Ile Tyr Tyr Val Gly Tyr Ser Gln Gly Thr Thr Met Gly 180 185 190 Phe Ile Ala Phe Ser Thr Met Pro Glu Leu Ala Gln Lys Ile Lys Met 195 200 205 Tyr Phe Ala Leu Ala Pro Ile Ala Thr Val Lys His Ala Lys Ser Pro 210 215 220 Gly Thr Lys Phe Leu Leu Leu Pro Asp Met Met Ile Lys Gly Leu Phe 225 230 235 240 Gly Lys Lys Glu Phe Leu Tyr Gln Thr Arg Phe Leu Arg Gln Leu Val 245 250 255 Ile Tyr Leu Cys Gly Gln Val Ile Leu Asp Gln Ile Cys Ser Asn Ile 260 265 270 Met Leu Leu Leu Gly Gly Phe Asn Thr Asn Asn Met Asn Met Ser Arg 275 280 285 Ala Ser Val Tyr Ala Ala His Thr Leu Ala Gly Thr Ser Val Gln Asn 290 295 300 Ile Leu His Trp Ser Gln Ala Val Asn Ser Gly Glu Leu Arg Ala Phe 305 310 315 320 Asp Trp Gly Ser Glu Thr Lys Asn Leu Glu Lys Cys Asn Gln Pro Thr 325 330 335 Pro Val Arg Tyr Arg Val Arg Asp Met Thr Val Pro Thr Ala Met Trp 340 345 350 Thr Gly Gly Gln Asp Trp Leu Ser Asn Pro Glu Asp Val Lys Met Leu 355 360 365 Leu Ser Glu Val Thr Asn Leu Ile Tyr His Lys Asn Ile Pro Glu Trp 370 375 380 Ala His Val Asp Phe Ile Trp Gly Leu Asp Ala Pro His Arg Met Tyr 385 390 395 400 Asn Glu Ile Ile His Leu Met Gln Gln Glu Glu Thr Asn Leu Ser Gln 405 410 415 Gly Arg Cys Glu Ala Val Leu 420 48 33 PRT Homo sapiens 48 Met Leu Glu Thr Leu Ser Arg Gln Trp Ile Val Ser His Arg Met Glu 1 5 10 15 Met Trp Leu Leu Ile Leu Val Ala Tyr Met Phe Gln Arg Asn Val Asn 20 25 30 Ser 49 390 PRT Homo sapiens 49 Val His Met Pro Thr Lys Ala Val Asp Pro Glu Ala Phe Met Asn Ile 1 5 10 15 Ser Glu Ile Ile Gln His Gln Gly Tyr Pro Cys Glu Glu Tyr Glu Val 20 25 30 Ala Thr Glu Asp Gly Tyr Ile Leu Ser Val Asn Arg Ile Pro Arg Gly 35 40 45 Leu Val Gln Pro Lys Lys Thr Gly Ser Arg Pro Val Val Leu Leu Gln 50 55 60 His Gly Leu Val Gly Gly Ala Ser Asn Trp Ile Ser Asn Leu Pro Asn 65 70 75 80 Asn Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly Phe Asp Val Trp Met 85 90 95 Gly Asn Ser Arg Gly Asn Ala Trp Ser Arg Lys His Lys Thr Leu Ser 100 105 110 Ile Asp Gln Asp Glu Phe Trp Ala Phe Ser Tyr Asp Glu Met Ala Arg 115 120 125 Phe Asp Leu Pro Ala Val Ile Asn Phe Ile Leu Gln Lys Thr Gly Gln 130 135 140 Glu Lys Ile Tyr Tyr Val Gly Tyr Ser Gln Gly Thr Thr Met Gly Phe 145 150 155 160 Ile Ala Phe Ser Thr Met Pro Glu Leu Ala Gln Lys Ile Lys Met Tyr 165 170 175 Phe Ala Leu Ala Pro Ile Ala Thr Val Lys His Ala Lys Ser Pro Gly 180 185 190 Thr Lys Phe Leu Leu Leu Pro Asp Met Met Ile Lys Gly Leu Phe Gly 195 200 205 Lys Lys Glu Phe Leu Tyr Gln Thr Arg Phe Leu Arg Gln Leu Val Ile 210 215 220 Tyr Leu Cys Gly Gln Val Ile Leu Asp Gln Ile Cys Ser Asn Ile Met 225 230 235 240 Leu Leu Leu Gly Gly Phe Asn Thr Asn Asn Met Asn Met Ser Arg Ala 245 250 255 Ser Val Tyr Ala Ala His Thr Leu Ala Gly Thr Ser Val Gln Asn Ile 260 265 270 Leu His Trp Ser Gln Ala Val Asn Ser Gly Glu Leu Arg Ala Phe Asp 275 280 285 Trp Gly Ser Glu Thr Lys Asn Leu Glu Lys Cys Asn Gln Pro Thr Pro 290 295 300 Val Arg Tyr Arg Val Arg Asp Met Thr Val Pro Thr Ala Met Trp Thr 305 310 315 320 Gly Gly Gln Asp Trp Leu Ser Asn Pro Glu Asp Val Lys Met Leu Leu 325 330 335 Ser Glu Val Thr Asn Leu Ile Tyr His Lys Asn Ile Pro Glu Trp Ala 340 345 350 His Val Asp Phe Ile Trp Gly Leu Asp Ala Pro His Arg Met Tyr Asn 355 360 365 Glu Ile Ile His Leu Met Gln Gln Glu Glu Thr Asn Leu Ser Gln Gly 370 375 380 Arg Cys Glu Ala Val Leu 385 390 50 221 PRT Homo sapiens 50 Val His Met Pro Thr Lys Ala Val Asp Pro Glu Ala Phe Met Asn Ile 1 5 10 15 Ser Glu Ile Ile Gln His Gln Gly Tyr Pro Cys Glu Glu Tyr Glu Val 20 25 30 Ala Thr Glu Asp Gly Tyr Ile Leu Ser Val Asn Arg Ile Pro Arg Gly 35 40 45 Leu Val Gln Pro Lys Lys Thr Gly Ser Arg Pro Val Val Leu Leu Gln 50 55 60 His Gly Leu Val Gly Gly Ala Ser Asn Trp Ile Ser Asn Leu Pro Asn 65 70 75 80 Asn Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly Phe Asp Val Trp Met 85 90 95 Gly Asn Ser Arg Gly Asn Ala Trp Ser Arg Lys His Lys Thr Leu Ser 100 105 110 Ile Asp Gln Asp Glu Phe Trp Ala Phe Ser Tyr Asp Glu Met Ala Arg 115 120 125 Phe Asp Leu Pro Ala Val Ile Asn Phe Ile Leu Gln Lys Thr Gly Gln 130 135 140 Glu Lys Ile Tyr Tyr Val Gly Tyr Ser Gln Gly Thr Thr Met Gly Phe 145 150 155 160 Ile Ala Phe Ser Thr Met Pro Glu Leu Ala Gln Lys Ile Lys Met Tyr 165 170 175 Phe Ala Leu Ala Pro Ile Ala Thr Val Lys His Ala Lys Ser Pro Gly 180 185 190 Thr Lys Phe Leu Leu Leu Pro Asp Met Met Ile Lys Gly Leu Phe Gly 195 200 205 Lys Lys Glu Phe Leu Tyr Gln Thr Arg Phe Leu Arg Gln 210 215 220 51 25 PRT Homo sapiens 51 Leu Val Ile Tyr Leu Cys Gly Gln Val Ile Leu Asp Gln Ile Cys Ser 1 5 10 15 Asn Ile Met Leu Leu Leu Gly Gly Phe 20 25 52 144 PRT Homo sapiens 52 Asn Thr Asn Asn Met Asn Met Ser Arg Ala Ser Val Tyr Ala Ala His 1 5 10 15 Thr Leu Ala Gly Thr Ser Val Gln Asn Ile Leu His Trp Ser Gln Ala 20 25 30 Val Asn Ser Gly Glu Leu Arg Ala Phe Asp Trp Gly Ser Glu Thr Lys 35 40 45 Asn Leu Glu Lys Cys Asn Gln Pro Thr Pro Val Arg Tyr Arg Val Arg 50 55 60 Asp Met Thr Val Pro Thr Ala Met Trp Thr Gly Gly Gln Asp Trp Leu 65 70 75 80 Ser Asn Pro Glu Asp Val Lys Met Leu Leu Ser Glu Val Thr Asn Leu 85 90 95 Ile Tyr His Lys Asn Ile Pro Glu Trp Ala His Val Asp Phe Ile Trp 100 105 110 Gly Leu Asp Ala Pro His Arg Met Tyr Asn Glu Ile Ile His Leu Met 115 120 125 Gln Gln Glu Glu Thr Asn Leu Ser Gln Gly Arg Cys Glu Ala Val Leu 130 135 140 53 2133 DNA Homo sapiens 53 gtcgacccac gcgtccacgg cgagggctcc cggggcgcag cattgccccc cctgcaccac 60 ctcaccaaga tggctacttt gggacacaca ttccccttct atgctggccc caagccaacc 120 ttcccgatgg acaccacttt ggccagcatc atcatgatct ttctgactgc actggccacg 180 ttcatcgtca tcctgcctgg cattcgggga aagacgaggc tgttctggct gcttcgggtg 240 gtgaccagct tattcatcgg ggctgcaatc ctggctgtga atttcagttc tgagtggtct 300 gtgggccagg tcagcaccaa cacatcatac aaggccttca gttctgagtg gatcagcgct 360 gatattgggc tgcaggtcgg gctgggtgga gtcaacatca cactcacagg gacccccgtg 420 cagcagctga atgagaccat caattacaac gaggagttca cctggcgcct gggtgagaac 480 tatgctgagg agtgtgcaaa ggctctggag aaggggctgc cagaccctgt gttgtaccta 540 gctgagaagt tcactccaag aagcccatgt ggcctatacc gccagtaccg cctggcggga 600 cactacacct cagccatgct atgggtggca ttcctctgct ggctgctggc caatgtgatg 660 ctctccatgc ctgtgctggt atatggtggc tacatgctat tggccacggg catcttccag 720 ctgttggctc tgctcttctt ctccatggcc acatcactca cctcaccctg tcccctgcac 780 ctgggcgctt ctgtgctgca tactcaccat gggcctgcct tctggatcac attgaccaca 840 ggactgctgt gtgtgctgct gggcctggct atggcggtgg cccacaggat gcagcctcac 900 aggctgaagg ctttcttcaa ccagagtgtg gatgaagacc ccatgctgga gtggagtcct 960 gaggaaggtg gactcctgag cccccgctac cggtccatgg ctgacagtcc caagtcccag 1020 gacattcccc tgtcagaggc ttcctccacc aaggcatact gtaaggaggc acaccccaaa 1080 gatcctgatt gtgctttata acattcctcc ccgtggaggc cacctggact tccagtctgg 1140 ctccaaacct cattggcgcc ccataaaacc agcagaactg ccctcagggt ggctgttacc 1200 agacacccag caccaatcta cagacggagt agaaaaagga ggctctatat actgatgtta 1260 aaaaacaaaa caaaacaaaa agccctaagg gactgaagag atgctgggcc tgtccataaa 1320 gcctgttgcc atgataaggc caagcagggg ctagcttatc tgcacagcaa cccagccttt 1380 ccgtgctgcc ttgcctcttc aagatgctat tcactgaaac ctaacttcac ccccataaca 1440 ccagcagggt gggggttaca tatgattctc ctatggtttc ctctcatccc tcggcacctc 1500 ttgttttcct ttttcctggg ttccttttgt tcttccttta cttctccagc ttgtgtggcc 1560 ttttggtaca atgaaagaca gcactggaaa ggaggggaaa ccaaacttct catcctaggt 1620 ctaacattaa ccaactatgc cacattctct ttgagcttca gttcccaaat ttgctacata 1680 agattgcaag acttgccaag aatcttggga tttatctttc tatgccttgc tgacacctac 1740 cttggccctc aaacaccacc tcacaagaag ccaggtggga agttagggaa tcaactccaa 1800 aacgctattc cttcccaccc cactcagctg ggctagctga gtggcatcca ggacggggga 1860 gtgggtgacc tgcctcatca ctgccaccta acgtccccct ggggtggttc agaaagatgc 1920 tagctctggt agggtccctc cggcctcact agagggcgcc cctattactc tggagtcgac 1980 gcagagaatc aggtttcaca gcactgcgga gagtgtacta ggctgtctcc agcccagcga 2040 agctcatgag gacgtgcgac cccggcgcgg agaagccatg aaaattaatg ggaaaaacag 2100 tttttaaaaa aaaaaaaaaa aaagggcggc cgc 2133 54 1029 DNA Homo sapiens 54 atggctactt tgggacacac attccccttc tatgctggcc ccaagccaac cttcccgatg 60 gacaccactt tggccagcat catcatgatc tttctgactg cactggccac gttcatcgtc 120 atcctgcctg gcattcgggg aaagacgagg ctgttctggc tgcttcgggt ggtgaccagc 180 ttattcatcg gggctgcaat cctggctgtg aatttcagtt ctgagtggtc tgtgggccag 240 gtcagcacca acacatcata caaggccttc agttctgagt ggatcagcgc tgatattggg 300 ctgcaggtcg ggctgggtgg agtcaacatc acactcacag ggacccccgt gcagcagctg 360 aatgagacca tcaattacaa cgaggagttc acctggcgcc tgggtgagaa ctatgctgag 420 gagtgtgcaa aggctctgga gaaggggctg ccagaccctg tgttgtacct agctgagaag 480 ttcactccaa gaagcccatg tggcctatac cgccagtacc gcctggcggg acactacacc 540 tcagccatgc tatgggtggc attcctctgc tggctgctgg ccaatgtgat gctctccatg 600 cctgtgctgg tatatggtgg ctacatgcta ttggccacgg gcatcttcca gctgttggct 660 ctgctcttct tctccatggc cacatcactc acctcaccct gtcccctgca cctgggcgct 720 tctgtgctgc atactcacca tgggcctgcc ttctggatca cattgaccac aggactgctg 780 tgtgtgctgc tgggcctggc tatggcggtg gcccacagga tgcagcctca caggctgaag 840 gctttcttca accagagtgt ggatgaagac cccatgctgg agtggagtcc tgaggaaggt 900 ggactcctga gcccccgcta ccggtccatg gctgacagtc ccaagtccca ggacattccc 960 ctgtcagagg cttcctccac caaggcatac tgtaaggagg cacaccccaa agatcctgat 1020 tgtgcttta 1029 55 343 PRT Homo sapiens 55 Met Ala Thr Leu Gly His Thr Phe Pro Phe Tyr Ala Gly Pro Lys Pro 1 5 10 15 Thr Phe Pro Met Asp Thr Thr Leu Ala Ser Ile Ile Met Ile Phe Leu 20 25 30 Thr Ala Leu Ala Thr Phe Ile Val Ile Leu Pro Gly Ile Arg Gly Lys 35 40 45 Thr Arg Leu Phe Trp Leu Leu Arg Val Val Thr Ser Leu Phe Ile Gly 50 55 60 Ala Ala Ile Leu Ala Val Asn Phe Ser Ser Glu Trp Ser Val Gly Gln 65 70 75 80 Val Ser Thr Asn Thr Ser Tyr Lys Ala Phe Ser Ser Glu Trp Ile Ser 85 90 95 Ala Asp Ile Gly Leu Gln Val Gly Leu Gly Gly Val Asn Ile Thr Leu 100 105 110 Thr Gly Thr Pro Val Gln Gln Leu Asn Glu Thr Ile Asn Tyr Asn Glu 115 120 125 Glu Phe Thr Trp Arg Leu Gly Glu Asn Tyr Ala Glu Glu Cys Ala Lys 130 135 140 Ala Leu Glu Lys Gly Leu Pro Asp Pro Val Leu Tyr Leu Ala Glu Lys 145 150 155 160 Phe Thr Pro Arg Ser Pro Cys Gly Leu Tyr Arg Gln Tyr Arg Leu Ala 165 170 175 Gly His Tyr Thr Ser Ala Met Leu Trp Val Ala Phe Leu Cys Trp Leu 180 185 190 Leu Ala Asn Val Met Leu Ser Met Pro Val Leu Val Tyr Gly Gly Tyr 195 200 205 Met Leu Leu Ala Thr Gly Ile Phe Gln Leu Leu Ala Leu Leu Phe Phe 210 215 220 Ser Met Ala Thr Ser Leu Thr Ser Pro Cys Pro Leu His Leu Gly Ala 225 230 235 240 Ser Val Leu His Thr His His Gly Pro Ala Phe Trp Ile Thr Leu Thr 245 250 255 Thr Gly Leu Leu Cys Val Leu Leu Gly Leu Ala Met Ala Val Ala His 260 265 270 Arg Met Gln Pro His Arg Leu Lys Ala Phe Phe Asn Gln Ser Val Asp 275 280 285 Glu Asp Pro Met Leu Glu Trp Ser Pro Glu Glu Gly Gly Leu Leu Ser 290 295 300 Pro Arg Tyr Arg Ser Met Ala Asp Ser Pro Lys Ser Gln Asp Ile Pro 305 310 315 320 Leu Ser Glu Ala Ser Ser Thr Lys Ala Tyr Cys Lys Glu Ala His Pro 325 330 335 Lys Asp Pro Asp Cys Ala Leu 340 56 23 PRT Homo sapiens 56 Met Ala Thr Leu Gly His Thr Phe Pro Phe Tyr Ala Gly Pro Lys Pro 1 5 10 15 Thr Phe Pro Met Asp Thr Thr 20 57 112 PRT Homo sapiens 57 Asn Phe Ser Ser Glu Trp Ser Val Gly Gln Val Ser Thr Asn Thr Ser 1 5 10 15 Tyr Lys Ala Phe Ser Ser Glu Trp Ile Ser Ala Asp Ile Gly Leu Gln 20 25 30 Val Gly Leu Gly Gly Val Asn Ile Thr Leu Thr Gly Thr Pro Val Gln 35 40 45 Gln Leu Asn Glu Thr Ile Asn Tyr Asn Glu Glu Phe Thr Trp Arg Leu 50 55 60 Gly Glu Asn Tyr Ala Glu Glu Cys Ala Lys Ala Leu Glu Lys Gly Leu 65 70 75 80 Pro Asp Pro Val Leu Tyr Leu Ala Glu Lys Phe Thr Pro Arg Ser Pro 85 90 95 Cys Gly Leu Tyr Arg Gln Tyr Arg Leu Ala Gly His Tyr Thr Ser Ala 100 105 110 58 22 PRT Homo sapiens 58 Thr Ser Leu Thr Ser Pro Cys Pro Leu His Leu Gly Ala Ser Val Leu 1 5 10 15 His Thr His His Gly Pro 20 59 19 PRT Homo sapiens 59 Leu Ala Ser Ile Ile Met Ile Phe Leu Thr Ala Leu Ala Thr Phe Ile 1 5 10 15 Val Ile Leu 60 20 PRT Homo sapiens 60 Leu Phe Trp Leu Leu Arg Val Val Thr Ser Leu Phe Ile Gly Ala Ala 1 5 10 15 Ile Leu Ala Val 20 61 22 PRT Homo sapiens 61 Met Leu Trp Val Ala Phe Leu Cys Trp Leu Leu Ala Asn Val Met Leu 1 5 10 15 Ser Met Pro Val Leu Val 20 62 17 PRT Homo sapiens 62 Leu Ala Thr Gly Ile Phe Gln Leu Leu Ala Leu Leu Phe Phe Ser Met 1 5 10 15 Ala 63 22 PRT Homo sapiens 63 Ala Phe Trp Ile Thr Leu Thr Thr Gly Leu Leu Cys Val Leu Leu Gly 1 5 10 15 Leu Ala Met Ala Val Ala 20 64 8 PRT Homo sapiens 64 Pro Gly Ile Arg Gly Lys Thr Arg 1 5 65 6 PRT Homo sapiens 65 Tyr Gly Gly Tyr Met Leu 1 5 66 72 PRT Homo sapiens 66 His Arg Met Gln Pro His Arg Leu Lys Ala Phe Phe Asn Gln Ser Val 1 5 10 15 Asp Glu Asp Pro Met Leu Glu Trp Ser Pro Glu Glu Gly Gly Leu Leu 20 25 30 Ser Pro Arg Tyr Arg Ser Met Ala Asp Ser Pro Lys Ser Gln Asp Ile 35 40 45 Pro Leu Ser Glu Ala Ser Ser Thr Lys Ala Tyr Cys Lys Glu Ala His 50 55 60 Pro Lys Asp Pro Asp Cys Ala Leu 65 70 67 4928 DNA Mus sp. 67 gtcgacccac gcgtccgccc ggctcccggt gctgccccct ctgccccggg ccgcgcccgg 60 gggtcccgca ctgacggccc atggcgccgc ccgccgcccg tctcgcgctg ctctccgccg 120 ctgcgctcac tctggcggcc cggcccgcgc ccggtccccg ctccggcccc gagtgcttca 180 cagccaacgg tgcagattac aggggaacac agagctggac agcgctgcaa ggtgggaagc 240 catgtctgtt ctggaacgag actttccagc atccgtacaa cacgctgaag taccccaacg 300 gggaaggagg actgggcgag cacaattatt gcagaaatcc agatggagac gtgagccctt 360 ggtgctacgt ggccgagcat gaggacggag tctactggaa gtactgtgaa attcctgcct 420 gccagatgcc tggaaacctt ggctgctaca aggatcatgg aaacccacct cctctcacgg 480 gcaccagtaa aacctctaac aagctcacca tacaaacctg tatcagcttc tgtcggagtc 540 agagattcaa gtttgctggg atggagtcag gctatgcctg cttctgtggg aacaatcctg 600 actactggaa gcacggggag gcggccagca ccgagtgcaa tagtgtctgc ttcggggacc 660 acacgcagcc ctgcggtggg gacggcagga ttatcctctt tgacactctc gtgggcgcct 720 gcggtgggaa ctactcagcc atggcagccg tggtgtactc ccctgacttc cctgacacct 780 acgccactgg cagagtctgc tactggacca tccgggttcc aggagcctct cgcatccatt 840 tcaacttcac cctgtttgat atcagggact ctgcagacat ggtggagctg ctggacggct 900 acacccaccg cgtcctggtc cggctcagtg ggaggagccg cccgcctctg tctttcaatg 960 tctctctgga ttttgtcatt ttgtatttct tctctgatcg catcaatcag gcccagggat 1020 ttgctgtgtt gtaccaagcc accaaggagg aaccgccaca ggagagacct gctgtcaacc 1080 agaccctggc agaggtgatc accgagcaag ccaacctcag tgtcagcgct gcccactcct 1140 ccaaagtcct ctatgtcatc acccccagcc ccagccaccc tccgcagact gcccaggtag 1200 ccattcctgg gcaccgtcag ttggggccaa cagccacaga gtggaaggat ggactgtgta 1260 cggcctggcg accctcctca tcctcacagt cacagcagtt gtcgcaaaga ttcttctgca 1320 tgtcacattt aaatctcatc gagtccctgc atcaggagac cttagggact gtcgtcagcc 1380 tggggcttct ggagatatct ggaccatttt ctatgaacct tccactacaa tctccatctt 1440 taagaagaag ctcaagggtc agagtcaaca agatgaccgc aatcccctcg tgagtgactg 1500 aagcccacgc ctgcatgaga ggctccgctc caagctcgag tttgctcccc tgagttctcc 1560 tctgatgagt tccctgcctt cccattcacc accatctctt ttgggagcac cctgctttag 1620 aggcagccca gcctgggatc ctccatcaca tgtaccagcc tggctgctct gctggggatg 1680 gtaagacagg cccaggctga caggacacag ctggacctga ctccagaaga ctcttgggtg 1740 gtggggaggt atagtgtagg atgagttttc ttgcttcttc tctgttttgt ccacatacag 1800 atcggtttcc cctgtcttta cagtttgcaa tagagccaga ctgaaagaac tgtcaggttt 1860 tctaggctgg cctggttccc cactaagagt ggcattggcg ccctagaggc ccagaggccc 1920 agtgtaggct tggagctttc tctgctgcca actaccatgt gtcatctagt ccgaggggac 1980 tgagagcagg gccacaccag atgtcatctt tctagagggt tctttttagt acccactgac 2040 caatggggca agcctgagga ttggtccatc tgtttgtcca tggaacagac acagtgaact 2100 tcctggatac tagacttaac tagcctagcc ctcaagtagt tgccaatcct gtggaatcag 2160 aattcagcct gtcttcctgt cctcagccca agcctgtagc ctagagctgg ggctgtagcc 2220 tagagctggg gctgtagcct agagctgggg ctgtagcaca gagctggggc tgtagcctag 2280 agctggggct gtagcacaga gctggggctg tagcctagag ctggggctgt agcacagagc 2340 tggggctgta gcacagagct ggggctgtag cctagagctg gggctgtagc acagagctgg 2400 ggctgtaact cagcgatcaa gagcttgctt tgtatacatc ggaccctagg ttctatccca 2460 gcactatcag aaggtgggag agaaaaagac tgcaccatag catgcgggca gcatctgtgg 2520 ttcctacgtg aggtgtcatc attttaaaag cagatcaaaa ctaccgcgag ttttgtcctt 2580 tgtcccttat catgggagca gagtaggagt aagggctctg gtcttgctca ttgtccccca 2640 gacagggagg caggaaaagg tcaggcttgg gaactggaga tcctcccagg aaaagctgca 2700 agattgagag acccagctgc agttgggaga ggaagggcca tccccgactg agaagtcctg 2760 cagtctggaa gtggcctttg tcagcagcag ctgtgccctg aaggtagacc ttggtcactc 2820 tcctgccagc ccttgagcct ctgctctcct gggtaccctc ctggaacacc atgctaacct 2880 tccccgagtc tctcagtcac tgccattgag gcctctcctc tagctgctgc tccccaggac 2940 tgtctggggc catctgggga tcagggagag gcagcaggag tactgacgag gcagtgacct 3000 gagctgatga gtcaaccaga ggacaccaga gtctacagtg ggctggctgc tggctcagct 3060 cctatgggag gcctacaggg gtactaagct agggggtcat catctcattt gatctgggaa 3120 aggctacagg ctcctggatg tgaagacagg cccactacat aagaagacca ctggaaatag 3180 actgacagga gcaggttcca ctctaggctg tccatagcgt ttgcaggact cccctgagac 3240 caagtgttga gtcacagagt gccatgtgcg tagtgcataa aggatatggg ttcttaacca 3300 gggaaggctc atagcaggcc aggacatttt ttcagctcag agcactggcc ccaggcttcc 3360 tctaagccac cactcacctg tctcttccta tctcggacac aggaagcaag ccccagtgtg 3420 gtggcagctg cggctcagca ttggtgtccc caggaagggc ggtggatgtg cccacgctcc 3480 ttttgctgtg ggcctggcac agcccaacac tgcagggccc accttctctc ttggggggta 3540 gggacacata aggaaaacta acccacctcc aacaacagca gaggacagtg ggaaggaagg 3600 gctgtaaatc acccaggcca gacctccaga aatgacaggc acagtctgtt agaacctgta 3660 ggcagccagt cacagagggc ctttgtgctg gtaacaccct gcctggagca taggggtaag 3720 ccgagggaga agagcagccc tcagagacat cagctaaaaa cataggtgcc ctatgtccct 3780 cccttcctgt cacactgctt acaaagcaga gacagagtag gaaagaggtc ttcatcctct 3840 cccacatcag caaggatagg gctgcggctg cctaaagtga gcaaggagaa cagagctctg 3900 gacttctcta aatgtgggct ctggcttcag actcctcagc caaaagctct tgaagatcaa 3960 agctctggcg ggtacagctg tcctggcctg tgggccagcc catgggatgt gcctgggcca 4020 ggtgccaccc cacggctcac tgtcatccca ggagggaccc cacctgatgc tcctcatcat 4080 ccgctggcct gacactatca gagctcgcgc cggctgttgc cagggacaga ctgactacac 4140 ttgaccttca agagcactta gaagtggatg gcctccagac tctgtcagcc tctgcagggg 4200 ccacacaagt ctcccgagcc aagtccacaa gcctccatgg ttccctggct cctctcctgt 4260 ggagtgtcct gtttgatgtc tgaggtctgc tttgggtacc gccctgggaa ctgctaacct 4320 ccgattggtc cctttgtgtc tctgtttact gtcctcttct acctccaggt cacttagctc 4380 tggctgctct ggctgggagt ggggggtggg gatgctggct gcacccccac cctggtctgc 4440 caacagaacc tgggggcctc acacgggctc ctgtcttgcc aagctggagc tgagcacact 4500 ggcccaggct gagtggggca gagcaaacaa gtggaagggg atctctctcc ttagagggag 4560 gtggccgaag gtgtagatcc agcgagggag ctgccatccc cgccaccttc atagcagcaa 4620 gaccttccca tttccaatct caccctccag cagggatatg actttggaca acaaggcttt 4680 atttgtaaat atgctcttaa tatgcaactt tgagaataag atagaaacat catgtatttt 4740 aaaatataaa atgaagtgtg acacactgta tacaatttaa tatatatttt taggattttg 4800 ttatttaaga aaatggaatg tgatggtact taacttttac aaaagagaga aaatgttatt 4860 tttactgttt gaagaaaata aatattctca ttgttgtaga aaaaaaaaaa aaaaaaaagg 4920 gcggccgc 4928 68 1410 DNA Mus sp. 68 atggcgccgc ccgccgcccg tctcgcgctg ctctccgccg ctgcgctcac tctggcggcc 60 cggcccgcgc ccggtccccg ctccggcccc gagtgcttca cagccaacgg tgcagattac 120 aggggaacac agagctggac agcgctgcaa ggtgggaagc catgtctgtt ctggaacgag 180 actttccagc atccgtacaa cacgctgaag taccccaacg gggaaggagg actgggcgag 240 cacaattatt gcagaaatcc agatggagac gtgagccctt ggtgctacgt ggccgagcat 300 gaggacggag tctactggaa gtactgtgaa attcctgcct gccagatgcc tggaaacctt 360 ggctgctaca aggatcatgg aaacccacct cctctcacgg gcaccagtaa aacctctaac 420 aagctcacca tacaaacctg tatcagcttc tgtcggagtc agagattcaa gtttgctggg 480 atggagtcag gctatgcctg cttctgtggg aacaatcctg actactggaa gcacggggag 540 gcggccagca ccgagtgcaa tagtgtctgc ttcggggacc acacgcagcc ctgcggtggg 600 gacggcagga ttatcctctt tgacactctc gtgggcgcct gcggtgggaa ctactcagcc 660 atggcagccg tggtgtactc ccctgacttc cctgacacct acgccactgg cagagtctgc 720 tactggacca tccgggttcc aggagcctct cgcatccatt tcaacttcac cctgtttgat 780 atcagggact ctgcagacat ggtggagctg ctggacggct acacccaccg cgtcctggtc 840 cggctcagtg ggaggagccg cccgcctctg tctttcaatg tctctctgga ttttgtcatt 900 ttgtatttct tctctgatcg catcaatcag gcccagggat ttgctgtgtt gtaccaagcc 960 accaaggagg aaccgccaca ggagagacct gctgtcaacc agaccctggc agaggtgatc 1020 accgagcaag ccaacctcag tgtcagcgct gcccactcct ccaaagtcct ctatgtcatc 1080 acccccagcc ccagccaccc tccgcagact gcccaggtag ccattcctgg gcaccgtcag 1140 ttggggccaa cagccacaga gtggaaggat ggactgtgta cggcctggcg accctcctca 1200 tcctcacagt cacagcagtt gtcgcaaaga ttcttctgca tgtcacattt aaatctcatc 1260 gagtccctgc atcaggagac cttagggact gtcgtcagcc tggggcttct ggagatatct 1320 ggaccatttt ctatgaacct tccactacaa tctccatctt taagaagaag ctcaagggtc 1380 agagtcaaca agatgaccgc aatcccctcg 1410 69 470 PRT Mus sp. 69 Met Ala Pro Pro Ala Ala Arg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala Ala Arg Pro Ala Pro Gly Pro Arg Ser Gly Pro Glu Cys 20 25 30 Phe Thr Ala Asn Gly Ala Asp Tyr Arg Gly Thr Gln Ser Trp Thr Ala 35 40 45 Leu Gln Gly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His 50 55 60 Pro Tyr Asn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu 65 70 75 80 His Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr 85 90 95 Val Ala Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro 100 105 110 Ala Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn 115 120 125 Pro Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile 130 135 140 Gln Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly 145 150 155 160 Met Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp Tyr Trp 165 170 175 Lys His Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys Phe Gly 180 185 190 Asp His Thr Gln Pro Cys Gly Gly Asp Gly Arg Ile Ile Leu Phe Asp 195 200 205 Thr Leu Val Gly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ala Ala Val 210 215 220 Val Tyr Ser Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg Val Cys 225 230 235 240 Tyr Trp Thr Ile Arg Val Pro Gly Ala Ser Arg Ile His Phe Asn Phe 245 250 255 Thr Leu Phe Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu Leu Asp 260 265 270 Gly Tyr Thr His Arg Val Leu Val Arg Leu Ser Gly Arg Ser Arg Pro 275 280 285 Pro Leu Ser Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe 290 295 300 Ser Asp Arg Ile Asn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala 305 310 315 320 Thr Lys Glu Glu Pro Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Leu 325 330 335 Ala Glu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala His 340 345 350 Ser Ser Lys Val Leu Tyr Val Ile Thr Pro Ser Pro Ser His Pro Pro 355 360 365 Gln Thr Ala Gln Val Ala Ile Pro Gly His Arg Gln Leu Gly Pro Thr 370 375 380 Ala Thr Glu Trp Lys Asp Gly Leu Cys Thr Ala Trp Arg Pro Ser Ser 385 390 395 400 Ser Ser Gln Ser Gln Gln Leu Ser Gln Arg Phe Phe Cys Met Ser His 405 410 415 Leu Asn Leu Ile Glu Ser Leu His Gln Glu Thr Leu Gly Thr Val Val 420 425 430 Ser Leu Gly Leu Leu Glu Ile Ser Gly Pro Phe Ser Met Asn Leu Pro 435 440 445 Leu Gln Ser Pro Ser Leu Arg Arg Ser Ser Arg Val Arg Val Asn Lys 450 455 460 Met Thr Ala Ile Pro Ser 465 470 70 760 PRT Mus sp. 70 Met Ala Leu Pro Ser Leu Gly Gln Asp Ser Trp Ser Leu Leu Arg Val 1 5 10 15 Phe Phe Phe Gln Leu Phe Leu Leu Pro Ser Leu Pro Pro Ala Ser Gly 20 25 30 Thr Gly Gly Gln Gly Pro Met Pro Arg Val Lys Tyr His Ala Gly Asp 35 40 45 Gly His Arg Ala Leu Ser Phe Phe Gln Gln Lys Gly Leu Arg Asp Phe 50 55 60 Asp Thr Leu Leu Leu Ser Asp Asp Gly Asn Thr Leu Tyr Val Gly Ala 65 70 75 80 Arg Glu Thr Val Leu Ala Leu Asn Ile Gln Asn Pro Gly Ile Pro Arg 85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Glu Arg Lys Lys Thr Glu 100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn Phe Ile 115 120 125 Arg Val Leu Val Ser Tyr Asn Ala Thr His Leu Tyr Ala Cys Gly Thr 130 135 140 Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser Leu 145 150 155 160 Leu Leu Pro Ile Leu Ile Asp Lys Val Met Asp Gly Lys Gly Gln Ser 165 170 175 Pro Leu Thr Leu Phe Thr Ser Thr Gln Ala Val Leu Val Asp Gly Met 180 185 190 Leu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile Leu 195 200 205 Met Arg Thr Leu Gly Ser His Pro Val Leu Lys Thr Asp Ile Phe Leu 210 215 220 Arg Trp Leu His Ala Asp Ala Ser Phe Val Ala Ala Ile Pro Ser Thr 225 230 235 240 Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245 250 255 Phe Glu Glu Leu Tyr Ile Ser Arg Val Ala Gln Val Cys Lys Asn Asp 260 265 270 Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu Lys 275 280 285 Ala Gln Leu Leu Cys Ala Gln Pro Gly Gln Leu Pro Phe Asn Ile Ile 290 295 300 Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Ser Val Ser Arg Ile 305 310 315 320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser Ser 325 330 335 Ala Val Cys Ala Phe Ser Leu Thr Asp Ile Glu Arg Val Phe Lys Gly 340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr Tyr Arg 355 360 365 Gly Ser Glu Val Ser Pro Arg Pro Gly Ser Cys Ser Met Gly Pro Ser 370 375 380 Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe Leu Met Asp Glu 385 390 395 400 His Val Val Gly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr Thr 405 410 415 Arg Leu Ala Val Glu Ser Ala Arg Gly Leu Asp Gly Ser Ser His Val 420 425 430 Val Met Tyr Leu Gly Thr Ser Thr Gly Pro Leu His Lys Ala Val Val 435 440 445 Pro Gln Asp Ser Ser Ala Tyr Leu Val Glu Glu Ile Gln Leu Ser Pro 450 455 460 Asp Ser Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Ala Gln Gly Ala 465 470 475 480 Val Phe Ala Gly Phe Ser Gly Gly Ile Trp Arg Val Pro Arg Ala Asn 485 490 495 Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro 500 505 510 His Cys Ala Trp Asp Pro Glu Ser Arg Leu Cys Ser Leu Leu Ser Gly 515 520 525 Ser Thr Lys Pro Trp Lys Gln Asp Met Glu Arg Gly Asn Pro Glu Trp 530 535 540 Val Cys Thr Arg Gly Pro Met Ala Arg Ser Pro Arg Arg Gln Ser Pro 545 550 555 560 Pro Gln Leu Ile Lys Glu Val Leu Thr Val Pro Asn Ser Ile Leu Glu 565 570 575 Leu Arg Cys Pro His Leu Ser Ala Leu Ala Ser Tyr His Trp Ser His 580 585 590 Gly Arg Ala Lys Ile Ser Glu Ala Ser Ala Thr Val Tyr Asn Gly Ser 595 600 605 Leu Leu Leu Leu Pro Gln Asp Gly Val Gly Gly Leu Tyr Gln Cys Val 610 615 620 Ala Thr Glu Asn Gly Tyr Ser Tyr Pro Val Val Ser Tyr Trp Val Asp 625 630 635 640 Ser Gln Asp Gln Pro Leu Ala Leu Asp Pro Glu Leu Ala Gly Val Pro 645 650 655 Arg Glu Arg Val Gln Val Pro Leu Thr Arg Val Gly Gly Gly Ala Ser 660 665 670 Met Ala Ala Gln Arg Ser Tyr Trp Pro His Phe Leu Ile Val Thr Val 675 680 685 Leu Leu Ala Ile Val Leu Leu Gly Val Leu Thr Leu Leu Leu Ala Ser 690 695 700 Pro Leu Gly Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Gly Met 705 710 715 720 Leu Pro Pro Arg Glu Lys Ala Pro Leu Ser Arg Asp Gln His Leu Gln 725 730 735 Pro Ser Lys Asp His Arg Thr Ser Ala Ser Asp Val Asp Ala Asp Asn 740 745 750 Asn His Leu Gly Ala Glu Val Ala 755 760 71 3046 DNA Mus sp. 71 ctcggacgcc tgggttaggg gtctgtactg ctggggaacc atctggtgac catctcaggc 60 tgaccatggc cctaccatcc ctgggccagg actcatggag tctcctgcgt gtttttttct 120 tccaactctt cctgctgcca tcactgccac ctgcttctgg gactggtggt caggggccca 180 tgcccagagt caaataccat gctggagacg ggcacagggc cctcagcttc ttccaacaaa 240 aaggcctccg agactttgac acgctgctcc tgagtgacga tggcaacact ctctatgtgg 300 gggctcgaga gaccgtcctg gccttgaata tccagaaccc aggaatccca aggctaaaga 360 acatgatacc ctggccagcc agtgagagaa aaaagaccga atgtgccttt aagaagaaga 420 gcaatgagac acagtgtttc aacttcattc gagtcctggt ctcttacaat gctactcacc 480 tctatgcctg tgggaccttt gccttcagcc ctgcctgtac cttcattgaa ctccaagatt 540 ccctcctgtt gcccatcttg atagacaagg tcatggacgg gaagggccaa agccctttga 600 ccctgttcac aagcacacaa gctgtcttgg tcgatgggat gctttattcc ggcaccatga 660 acaacttcct gggcagcgag cccatcctga tgcggacact gggatcccat cctgttctca 720 agactgacat cttcttacgc tggctgcacg cggatgcctc cttcgtggca gccattccat 780 ccacccaggt cgtctatttc ttctttgagg agacagccag cgagtttgac ttctttgaag 840 agctgtatat atccagggtg gctcaagtct gcaagaacga cgtgggcggt gaaaagctgc 900 tgcagaagaa gtggaccacc ttcctcaaag cccagttgct ctgcgctcag ccagggcagc 960 tgccattcaa catcatccgc cacgcggtcc tgctgcccgc cgattctccc tctgtttccc 1020 gcatctacgc agtctttacc tcccagtggc aggttggcgg gaccaggagc tcagcagtct 1080 gtgccttctc tctcacggac attgagcgag tctttaaagg gaagtacaag gagctgaaca 1140 aggagacctc ccgctggacc acttaccggg gctcagaggt cagcccgagg ccaggcagtt 1200 gctccatggg cccctcctct gacaaagcct tgaccttcat gaaggaccat tttctgatgg 1260 atgagcacgt ggtaggaaca cccctgctgg tgaagtctgg tgtggagtac acacggcttg 1320 ctgtggagtc agctcggggc cttgatggga gcagccatgt ggtcatgtat ctgggtacct 1380 ccacgggtcc cctgcacaag gctgtggtgc ctcaggacag cagtgcttat ctcgtggagg 1440 agattcagct gagccctgac tctgagcctg ttcgaaacct gcagctggcc cccgcccagg 1500 gtgcagtgtt tgcaggcttc tctggaggca tctggagagt tcccagggcc aattgcagtg 1560 tctacgagag ctgtgtggac tgtgtgcttg ccagggaccc tcactgtgcc tgggaccctg 1620 aatcaagact ctgcagcctt ctgtctggct ctaccaagcc ttggaagcag gacatggaac 1680 gcggcaaccc ggagtgggta tgcacccgtg gccccatggc caggagcccc cggcgtcaga 1740 gcccccctca actaattaaa gaagtcctga cagtccccaa ctccatcctg gagctgcgct 1800 gcccccacct gtcagcactg gcctcttacc actggagtca tggccgagcc aaaatctcag 1860 aagcctctgc taccgtctac aatggctccc tcttgctgct gccgcaggat ggtgtcgggg 1920 gcctctacca gtgtgtggcg actgagaacg gctactcata ccctgtggtc tcctattggg 1980 tagacagcca ggaccagccc ctggcgctgg accctgagct ggcgggcgtt ccccgtgagc 2040 gtgtgcaggt cccgctgacc agggtcggag gcggagcttc catggctgcc cagcggtcct 2100 actggcccca ttttctcatc gttaccgtcc tcctggccat cgtgctcctg ggagtgctca 2160 ctctcctcct cgcttcccca ctgggggcgc tgcgggctcg gggtaaggtt cagggctgtg 2220 ggatgctgcc ccccagggaa aaggctccac tgagcaggga ccagcacctc cagccctcca 2280 aggaccacag gacctctgcc agtgacgtag atgccgacaa caaccatctg ggcgccgaag 2340 tggcttaaac agggacacag atccgcagct gagcagagca agccactggc cttgttggct 2400 atgccaggca cagtgccact ctgaccaggg taggaggctc tcctgctaac gtgtgtcacc 2460 tacagcaccc agtaggtcct cccctgtggg actctcttct gcaagcacat tgggctgtct 2520 ccatacctgt acttgtgctg tgacaggaag agccagacag gtttctttga ttttgattga 2580 cccaagagcc ctgcctgtaa caaacgtgct ccaggagacc atgaaaggtg tggctgtctg 2640 ggattctgtg gtgacaaacc taagcatccg agcaagctgg ggctattcct gcaaactcca 2700 tcctgaacgc tgtcactcta gaagcagctg ctgctttgaa caccagccca ccctccttcc 2760 caagagtctc tatggagttg gccccttgtg tttcctttac cagtcgggcc atactgtttg 2820 ggaagtcatc tctgaagtct aaccaccttc cttcttggtt cagtttggac agattgttat 2880 tattgtctct gccctggcta gaatgggggc ataatctgag ccttgttccc ttgtccagtg 2940 tggctgaccc ttgacctctt ccttcctcct ccctttgttt tgggattcag aaaactgctt 3000 gtcacagaca atttattttt tattaaaaaa gatataagct ttaaag 3046 72 2915 DNA Mus sp. 72 gtcgacccac gcgtccggcc gcgcgtcctt ctgccggctt cagctcgtat ccccggagtc 60 cacccgcccg tcccggggtg cggactggcc ctgagctggc cgtacagccc ggcttcggac 120 ggtcctcgct ggagccatgg gccgccggct cggcagggtg gcggcgctgc tgctcgggct 180 gctagtggag tgcactgagg ccaaaaaaca ttgctggtat tttgaaggac tctatcccac 240 atactatata tgccgttcct atgaagactg ctgtggctcc aggtgctgtg tgagggccct 300 ttccatacag aggctgtggt atttttggtt cctgctgatg atgggtgtgc tgttctgctg 360 tggtgccggt ttcttcattc gccggcgcat gtatccgcca ccactcattg aggagcccac 420 attcaatgtg tcctatacca ggcagccacc aaatcctgct ccaggagcac agcaaatggg 480 accgccatat tacaccgacc ctggaggacc cgggatgaat cctgttggca ataccatggc 540 tatggctttc caggtccagc ccaattcacc tcacggaggc acaacttacc caccccctcc 600 ttcctactgc aacacgcctc caccccccta tgaacaggtg gtgaaggaca agtagcaaga 660 tgctacatca aaggcaaaga ggatggacag gcccttttgt ttaccttccc atcctcaccg 720 atacttgctg atagggtggt ccaagggaaa acttggatat tctcaaagca agcccagctc 780 tctttcaagt cttttgtgga ggacatttga atccacactg tctcctctgt tgcttctgtt 840 tctgatgtag tctgtgctct ctgagagagt gtggcaacag tccctgaggg ttgatattcc 900 tagggtgtcc agggtagatc ctcgggagag aggctaaggg gaaaggaagg catagcctgt 960 gtgttagggg gcagataaag tggtcaggct gagataagac tcacatgatg cagtagttgg 1020 cagtgaactt cgaagagaca ctatccacca tcccagccca ttctcctaat agaagctgtg 1080 gggctgtgtt gttgatgctc tttggtctcc actcacattt tgaaaatagg ctttcctctg 1140 caggaatagg aaagacccaa gtacatattt gcttccactt aaaaatgagg gtcagaacca 1200 ggcctcagtt ggacatctat agttaaataa aggccattag agaggggaaa tctttaagtt 1260 aggggaaatt ctctaaatgg agacattgcg ttttatgaat catcgtctgg cttttctttt 1320 agtgcatgta ttgaagtgag ggtgtccttt gagatcagat ggggagagtg aactctgcgg 1380 ggggtggggt gtctctactc agagggctcc aacacccttt tcttaggtag ttctggtgat 1440 gggttttatg ggcactatag agctgagggg cacattaggc cgggtagtta cattgaccct 1500 tggagaggaa gaggacagcc aaagaaactc agcaaagcaa gaccagcatt gctgagttag 1560 agctagggtt gtatgtgatc ccaacagaga tgtgctggcc tcagaagagg ggacgtttgt 1620 ggatagagcc gtgaaaacct acttagttgc acagatgaca taatcaaaag tagagaaaga 1680 agtgtagtta gagatgccat ttcccaggtg agaatcagag ctcatccata gatttacaag 1740 tagtggctgg agttaacagt atggagttct tttcccttgc gtagttagtc acgttgatgt 1800 gtatttaaac ccaggttgag accttgtgta ctaagagcaa ggaagtatag ctaagatgtc 1860 tagattattt atatgtagta tggtggggag tggggctgca aggaaggggg ctgacattgt 1920 aaatgagaaa atcagagcca tttgataaac tgttacttgt tggatcaggc atccaaaagt 1980 gtctcttgag tggacattga gtattcttta ccacctacaa gaccaggagg catggtgtca 2040 ttctccattg gggtatttat atgaggtaga ggttcaggaa tcgacagtag ctgtgtgggc 2100 ttagtttaag gactgaaagc atagggactg gtagacagtt tcataggaaa ctgcggggaa 2160 ggaatggata cctttaaaga cagtttgtgg atgcagatgc tgccacccat cattgagcac 2220 ccttgtgtct ctggcttcct gtcactggat ccagtacccc tccatgcttg ggtccttgtt 2280 ttacataaga caacaaagca caatgtctgc tgtttacaat caagacgact acatggtcca 2340 aacatttctt ctctcttcta tcacttgtgg ctttaacttc catttcctcc gttccttttt 2400 aaaatcaaga agcacagtca gagctgcccc tgggattgca tcagggaacg gctgatcaag 2460 gcattcagtg tccatgacta aatcttatct ttttgatagc aaatcctttt aagaaactga 2520 acaattgcta aggctcagca attttatact ccaatgtctg tgtaaggtaa attttgtttg 2580 ccattgagcc cacattggaa ttccttctga cgtcaacact gacaatgcct atggaaattg 2640 cacttctggg tatatgtccc agcatccttg ttttcttatg tttggtgagt aaggctcacc 2700 ccttccagca gctctacttc tgtgtgctga ggtcctgtag agccggggct tgggcacaga 2760 catgaggcag acttgtgcat gctctttctt ggcaacactt ggctcatatt tcttgttctc 2820 ttttgataga gtcctgtttc ctatgtattt aaaaaataat aaaagtgaat ttagtcaaaa 2880 aaaaaaaaaa aaaaaaaaaa aaaaagggcg gccgc 2915 73 516 DNA Mus sp. 73 atgggccgcc ggctcggcag ggtggcggcg ctgctgctcg ggctgctagt ggagtgcact 60 gaggccaaaa aacattgctg gtattttgaa ggactctatc ccacatacta tatatgccgt 120 tcctatgaag actgctgtgg ctccaggtgc tgtgtgaggg ccctttccat acagaggctg 180 tggtattttt ggttcctgct gatgatgggt gtgctgttct gctgtggtgc cggtttcttc 240 attcgccggc gcatgtatcc gccaccactc attgaggagc ccacattcaa tgtgtcctat 300 accaggcagc caccaaatcc tgctccagga gcacagcaaa tgggaccgcc atattacacc 360 gaccctggag gacccgggat gaatcctgtt ggcaatacca tggctatggc tttccaggtc 420 cagcccaatt cacctcacgg aggcacaact tacccacccc ctccttccta ctgcaacacg 480 cctccacccc cctatgaaca ggtggtgaag gacaag 516 74 172 PRT Mus sp. 74 Met Gly Arg Arg Leu Gly Arg Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Val Glu Cys Thr Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30 Tyr Pro Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45 Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp 50 55 60 Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly Ala Gly Phe Phe 65 70 75 80 Ile Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro Thr Phe 85 90 95 Asn Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln 100 105 110 Gln Met Gly Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro Val Gly Asn Thr Met Ala Met Ala Phe Gln Val Gln Pro Asn Ser 130 135 140 Pro His Gly Gly Thr Thr Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr 145 150 155 160 Pro Pro Pro Pro Tyr Glu Gln Val Val Lys Asp Lys 165 170 75 398 PRT Homo sapiens 75 Met Trp Leu Leu Leu Thr Met Ala Ser Leu Ile Ser Val Leu Gly Thr 1 5 10 15 Thr His Gly Leu Phe Gly Lys Leu His Pro Gly Ser Pro Glu Val Thr 20 25 30 Met Asn Ile Ser Gln Met Ile Thr Tyr Trp Gly Tyr Pro Asn Glu Glu 35 40 45 Tyr Glu Val Val Thr Glu Asp Gly Tyr Ile Leu Glu Val Asn Arg Ile 50 55 60 Pro Tyr Gly Lys Lys Asn Ser Gly Asn Thr Gly Gln Arg Pro Val Val 65 70 75 80 Phe Leu Gln His Gly Leu Leu Ala Ser Ala Thr Asn Trp Ile Ser Asn 85 90 95 Leu Pro Asn Asn Ser Leu Ala Phe Ile Leu Ala Asp Ala Gly Tyr Asp 100 105 110 Val Trp Leu Gly Asn Ser Arg Gly Asn Thr Trp Ala Arg Arg Asn Leu 115 120 125 Tyr Tyr Ser Pro Asp Ser Val Glu Phe Trp Ala Phe Ser Phe Asp Glu 130 135 140 Met Ala Lys Tyr Asp Leu Pro Ala Thr Ile Asp Phe Ile Val Lys Lys 145 150 155 160 Thr Gly Gln Lys Gln Leu His Tyr Val Gly His Ser Gln Gly Thr Thr 165 170 175 Ile Gly Phe Ile Ala Phe Ser Thr Asn Pro Ser Leu Ala Lys Arg Ile 180 185 190 Lys Thr Phe Tyr Ala Leu Ala Pro Val Ala Thr Val Lys Tyr Thr Lys 195 200 205 Ser Leu Ile Asn Lys Leu Arg Phe Val Pro Gln Ser Leu Phe Lys Phe 210 215 220 Ile Phe Gly Asp Lys Ile Phe Tyr Pro His Asn Phe Phe Asp Gln Phe 225 230 235 240 Leu Ala Thr Glu Val Cys Ser Arg Glu Met Leu Asn Leu Leu Cys Ser 245 250 255 Asn Ala Leu Phe Ile Ile Cys Gly Phe Asp Ser Lys Asn Phe Asn Thr 260 265 270 Ser Arg Leu Asp Val Tyr Leu Ser His Asn Pro Ala Gly Thr Ser Val 275 280 285 Gln Asn Met Phe His Trp Thr Gln Ala Val Lys Ser Gly Lys Phe Gln 290 295 300 Ala Tyr Asp Trp Gly Ser Pro Val Gln Asn Arg Met His Tyr Asp Gln 305 310 315 320 Ser Gln Pro Pro Tyr Tyr Asn Val Thr Ala Met Asn Val Pro Ile Ala 325 330 335 Val Trp Asn Gly Gly Lys Asp Leu Leu Ala Asp Pro Gln Asp Val Gly 340 345 350 Leu Leu Leu Pro Lys Leu Pro Asn Leu Ile Tyr His Lys Glu Ile Pro 355 360 365 Phe Tyr Asn His Leu Asp Phe Ile Trp Ala Met Asp Ala Pro Gln Glu 370 375 380 Val Tyr Asn Asp Ile Val Ser Met Ile Ser Glu Asp Lys Lys 385 390 395 76 760 PRT Mus sp. 76 Met Ala Leu Pro Ser Leu Gly Gln Asp Ser Trp Ser Leu Leu Arg Val 1 5 10 15 Phe Phe Phe Gln Leu Phe Leu Leu Pro Ser Leu Pro Pro Ala Ser Gly 20 25 30 Thr Gly Gly Gln Gly Pro Met Pro Arg Val Lys Tyr His Ala Gly Asp 35 40 45 Gly His Arg Ala Leu Ser Phe Phe Gln Gln Lys Gly Leu Arg Asp Phe 50 55 60 Asp Thr Leu Leu Leu Ser Asp Asp Gly Asn Thr Leu Tyr Val Gly Ala 65 70 75 80 Arg Glu Thr Val Leu Ala Leu Asn Ile Gln Asn Pro Gly Ile Pro Arg 85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Glu Arg Lys Lys Thr Glu 100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn Phe Ile 115 120 125 Arg Val Leu Val Ser Tyr Asn Ala Thr His Leu Tyr Ala Cys Gly Thr 130 135 140 Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser Leu 145 150 155 160 Leu Leu Pro Ile Leu Ile Asp Lys Val Met Asp Gly Lys Gly Gln Ser 165 170 175 Pro Leu Thr Leu Phe Thr Ser Thr Gln Ala Val Leu Val Asp Gly Met 180 185 190 Leu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile Leu 195 200 205 Met Arg Thr Leu Gly Ser His Pro Val Leu Lys Thr Asp Ile Phe Leu 210 215 220 Arg Trp Leu His Ala Asp Ala Ser Phe Val Ala Ala Ile Pro Ser Thr 225 230 235 240 Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245 250 255 Phe Glu Glu Leu Tyr Ile Ser Arg Val Ala Gln Val Cys Lys Asn Asp 260 265 270 Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu Lys 275 280 285 Ala Gln Leu Leu Cys Ala Gln Pro Gly Gln Leu Pro Phe Asn Ile Ile 290 295 300 Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Ser Val Ser Arg Ile 305 310 315 320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser Ser 325 330 335 Ala Val Cys Ala Phe Ser Leu Thr Asp Ile Glu Arg Val Phe Lys Gly 340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr Tyr Arg 355 360 365 Gly Ser Glu Val Ser Pro Arg Pro Gly Ser Cys Ser Met Gly Pro Ser 370 375 380 Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe Leu Met Asp Glu 385 390 395 400 His Val Val Gly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr Thr 405 410 415 Arg Leu Ala Val Glu Ser Ala Arg Gly Leu Asp Gly Ser Ser His Val 420 425 430 Val Met Tyr Leu Gly Thr Ser Thr Gly Pro Leu His Lys Ala Val Val 435 440 445 Pro Gln Asp Ser Ser Ala Tyr Leu Val Glu Glu Ile Gln Leu Ser Pro 450 455 460 Asp Ser Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Ala Gln Gly Ala 465 470 475 480 Val Phe Ala Gly Phe Ser Gly Gly Ile Trp Arg Val Pro Arg Ala Asn 485 490 495 Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro 500 505 510 His Cys Ala Trp Asp Pro Glu Ser Arg Leu Cys Ser Leu Leu Ser Gly 515 520 525 Ser Thr Lys Pro Trp Lys Gln Asp Met Glu Arg Gly Asn Pro Glu Trp 530 535 540 Val Cys Thr Arg Gly Pro Met Ala Arg Ser Pro Arg Arg Gln Ser Pro 545 550 555 560 Pro Gln Leu Ile Lys Glu Val Leu Thr Val Pro Asn Ser Ile Leu Glu 565 570 575 Leu Arg Cys Pro His Leu Ser Ala Leu Ala Ser Tyr His Trp Ser His 580 585 590 Gly Arg Ala Lys Ile Ser Glu Ala Ser Ala Thr Val Tyr Asn Gly Ser 595 600 605 Leu Leu Leu Leu Pro Gln Asp Gly Val Gly Gly Leu Tyr Gln Cys Val 610 615 620 Ala Thr Glu Asn Gly Tyr Ser Tyr Pro Val Val Ser Tyr Trp Val Asp 625 630 635 640 Ser Gln Asp Gln Pro Leu Ala Leu Asp Pro Glu Leu Ala Gly Val Pro 645 650 655 Arg Glu Arg Val Gln Val Pro Leu Thr Arg Val Gly Gly Gly Ala Ser 660 665 670 Met Ala Ala Gln Arg Ser Tyr Trp Pro His Phe Leu Ile Val Thr Val 675 680 685 Leu Leu Ala Ile Val Leu Leu Gly Val Leu Thr Leu Leu Leu Ala Ser 690 695 700 Pro Leu Gly Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Gly Met 705 710 715 720 Leu Pro Pro Arg Glu Lys Ala Pro Leu Ser Arg Asp Gln His Leu Gln 725 730 735 Pro Ser Lys Asp His Arg Thr Ser Ala Ser Asp Val Asp Ala Asp Asn 740 745 750 Asn His Leu Gly Ala Glu Val Ala 755 760 77 3046 DNA Mus sp. 77 ctcggacgcc tgggttaggg gtctgtactg ctggggaacc atctggtgac catctcaggc 60 tgaccatggc cctaccatcc ctgggccagg actcatggag tctcctgcgt gtttttttct 120 tccaactctt cctgctgcca tcactgccac ctgcttctgg gactggtggt caggggccca 180 tgcccagagt caaataccat gctggagacg ggcacagggc cctcagcttc ttccaacaaa 240 aaggcctccg agactttgac acgctgctcc tgagtgacga tggcaacact ctctatgtgg 300 gggctcgaga gaccgtcctg gccttgaata tccagaaccc aggaatccca aggctaaaga 360 acatgatacc ctggccagcc agtgagagaa aaaagaccga atgtgccttt aagaagaaga 420 gcaatgagac acagtgtttc aacttcattc gagtcctggt ctcttacaat gctactcacc 480 tctatgcctg tgggaccttt gccttcagcc ctgcctgtac cttcattgaa ctccaagatt 540 ccctcctgtt gcccatcttg atagacaagg tcatggacgg gaagggccaa agccctttga 600 ccctgttcac aagcacacaa gctgtcttgg tcgatgggat gctttattcc ggcaccatga 660 acaacttcct gggcagcgag cccatcctga tgcggacact gggatcccat cctgttctca 720 agactgacat cttcttacgc tggctgcacg cggatgcctc cttcgtggca gccattccat 780 ccacccaggt cgtctatttc ttctttgagg agacagccag cgagtttgac ttctttgaag 840 agctgtatat atccagggtg gctcaagtct gcaagaacga cgtgggcggt gaaaagctgc 900 tgcagaagaa gtggaccacc ttcctcaaag cccagttgct ctgcgctcag ccagggcagc 960 tgccattcaa catcatccgc cacgcggtcc tgctgcccgc cgattctccc tctgtttccc 1020 gcatctacgc agtctttacc tcccagtggc aggttggcgg gaccaggagc tcagcagtct 1080 gtgccttctc tctcacggac attgagcgag tctttaaagg gaagtacaag gagctgaaca 1140 aggagacctc ccgctggacc acttaccggg gctcagaggt cagcccgagg ccaggcagtt 1200 gctccatggg cccctcctct gacaaagcct tgaccttcat gaaggaccat tttctgatgg 1260 atgagcacgt ggtaggaaca cccctgctgg tgaagtctgg tgtggagtac acacggcttg 1320 ctgtggagtc agctcggggc cttgatggga gcagccatgt ggtcatgtat ctgggtacct 1380 ccacgggtcc cctgcacaag gctgtggtgc ctcaggacag cagtgcttat ctcgtggagg 1440 agattcagct gagccctgac tctgagcctg ttcgaaacct gcagctggcc cccgcccagg 1500 gtgcagtgtt tgcaggcttc tctggaggca tctggagagt tcccagggcc aattgcagtg 1560 tctacgagag ctgtgtggac tgtgtgcttg ccagggaccc tcactgtgcc tgggaccctg 1620 aatcaagact ctgcagcctt ctgtctggct ctaccaagcc ttggaagcag gacatggaac 1680 gcggcaaccc ggagtgggta tgcacccgtg gccccatggc caggagcccc cggcgtcaga 1740 gcccccctca actaattaaa gaagtcctga cagtccccaa ctccatcctg gagctgcgct 1800 gcccccacct gtcagcactg gcctcttacc actggagtca tggccgagcc aaaatctcag 1860 aagcctctgc taccgtctac aatggctccc tcttgctgct gccgcaggat ggtgtcgggg 1920 gcctctacca gtgtgtggcg actgagaacg gctactcata ccctgtggtc tcctattggg 1980 tagacagcca ggaccagccc ctggcgctgg accctgagct ggcgggcgtt ccccgtgagc 2040 gtgtgcaggt cccgctgacc agggtcggag gcggagcttc catggctgcc cagcggtcct 2100 actggcccca ttttctcatc gttaccgtcc tcctggccat cgtgctcctg ggagtgctca 2160 ctctcctcct cgcttcccca ctgggggcgc tgcgggctcg gggtaaggtt cagggctgtg 2220 ggatgctgcc ccccagggaa aaggctccac tgagcaggga ccagcacctc cagccctcca 2280 aggaccacag gacctctgcc agtgacgtag atgccgacaa caaccatctg ggcgccgaag 2340 tggcttaaac agggacacag atccgcagct gagcagagca agccactggc cttgttggct 2400 atgccaggca cagtgccact ctgaccaggg taggaggctc tcctgctaac gtgtgtcacc 2460 tacagcaccc agtaggtcct cccctgtggg actctcttct gcaagcacat tgggctgtct 2520 ccatacctgt acttgtgctg tgacaggaag agccagacag gtttctttga ttttgattga 2580 cccaagagcc ctgcctgtaa caaacgtgct ccaggagacc atgaaaggtg tggctgtctg 2640 ggattctgtg gtgacaaacc taagcatccg agcaagctgg ggctattcct gcaaactcca 2700 tcctgaacgc tgtcactcta gaagcagctg ctgctttgaa caccagccca ccctccttcc 2760 caagagtctc tatggagttg gccccttgtg tttcctttac cagtcgggcc atactgtttg 2820 ggaagtcatc tctgaagtct aaccaccttc cttcttggtt cagtttggac agattgttat 2880 tattgtctct gccctggcta gaatgggggc ataatctgag ccttgttccc ttgtccagtg 2940 tggctgaccc ttgacctctt ccttcctcct ccctttgttt tgggattcag aaaactgctt 3000 gtcacagaca atttattttt tattaaaaaa gatataagct ttaaag 3046 78 1436 PRT Bos sp. 78 Met Ala Leu Gly Arg His Leu Ser Leu Arg Gly Leu Cys Val Leu Leu 1 5 10 15 Leu Gly Thr Met Val Gly Gly Gln Ala Leu Glu Leu Arg Leu Lys Asp 20 25 30 Gly Val His Arg Cys Glu Gly Arg Val Glu Val Lys His Gln Gly Glu 35 40 45 Trp Gly Thr Val Asp Gly Tyr Arg Trp Thr Leu Lys Asp Ala Ser Val 50 55 60 Val Cys Arg Gln Leu Gly Cys Gly Ala Ala Ile Gly Phe Pro Gly Gly 65 70 75 80 Ala Tyr Phe Gly Pro Gly Leu Gly Pro Ile Trp Leu Leu Tyr Thr Ser 85 90 95 Cys Glu Gly Thr Glu Ser Thr Val Ser Asp Cys Glu His Ser Asn Ile 100 105 110 Lys Asp Tyr Arg Asn Asp Gly Tyr Asn His Gly Arg Asp Ala Gly Val 115 120 125 Val Cys Ser Gly Phe Val Arg Leu Ala Gly Gly Asp Gly Pro Cys Ser 130 135 140 Gly Arg Val Glu Val His Ser Gly Glu Ala Trp Ile Pro Val Ser Asp 145 150 155 160 Gly Asn Phe Thr Leu Ala Thr Ala Gln Ile Ile Cys Ala Glu Leu Gly 165 170 175 Cys Gly Lys Ala Val Ser Val Leu Gly His Glu Leu Phe Arg Glu Ser 180 185 190 Ser Ala Gln Val Trp Ala Glu Glu Phe Arg Cys Glu Gly Glu Glu Pro 195 200 205 Glu Leu Trp Val Cys Pro Arg Val Pro Cys Pro Gly Gly Thr Cys His 210 215 220 His Ser Gly Ser Ala Gln Val Val Cys Ser Ala Tyr Ser Glu Val Arg 225 230 235 240 Leu Met Thr Asn Gly Ser Ser Gln Cys Glu Gly Gln Val Glu Met Asn 245 250 255 Ile Ser Gly Gln Trp Arg Ala Leu Cys Ala Ser His Trp Ser Leu Ala 260 265 270 Asn Ala Asn Val Ile Cys Arg Gln Leu Gly Cys Gly Val Ala Ile Ser 275 280 285 Thr Pro Gly Gly Pro His Leu Val Glu Glu Gly Asp Gln Ile Leu Thr 290 295 300 Ala Arg Phe His Cys Ser Gly Ala Glu Ser Phe Leu Trp Ser Cys Pro 305 310 315 320 Val Thr Ala Leu Gly Gly Pro Asp Cys Ser His Gly Asn Thr Ala Ser 325 330 335 Val Ile Cys Ser Gly Asn Gln Ile Gln Val Leu Pro Gln Cys Asn Asp 340 345 350 Ser Val Ser Gln Pro Thr Gly Ser Ala Ala Ser Glu Asp Ser Ala Pro 355 360 365 Tyr Cys Ser Asp Ser Arg Gln Leu Arg Leu Val Asp Gly Gly Gly Pro 370 375 380 Cys Ala Gly Arg Val Glu Ile Leu Asp Gln Gly Ser Trp Gly Thr Ile 385 390 395 400 Cys Asp Asp Gly Trp Asp Leu Asp Asp Ala Arg Val Val Cys Arg Gln 405 410 415 Leu Gly Cys Gly Glu Ala Leu Asn Ala Thr Gly Ser Ala His Phe Gly 420 425 430 Ala Gly Ser Gly Pro Ile Trp Leu Asp Asn Leu Asn Cys Thr Gly Lys 435 440 445 Glu Ser His Val Trp Arg Cys Pro Ser Arg Gly Trp Gly Gln His Asn 450 455 460 Cys Arg His Lys Gln Asp Ala Gly Val Ile Cys Ser Glu Phe Leu Ala 465 470 475 480 Leu Arg Met Val Ser Glu Asp Gln Gln Cys Ala Gly Trp Leu Glu Val 485 490 495 Phe Tyr Asn Gly Thr Trp Gly Ser Val Cys Arg Asn Pro Met Glu Asp 500 505 510 Ile Thr Val Ser Thr Ile Cys Arg Gln Leu Gly Cys Gly Asp Ser Gly 515 520 525 Thr Leu Asn Ser Ser Val Ala Leu Arg Glu Gly Phe Arg Pro Gln Trp 530 535 540 Val Asp Arg Ile Gln Cys Arg Lys Thr Asp Thr Ser Leu Trp Gln Cys 545 550 555 560 Pro Ser Asp Pro Trp Asn Tyr Asn Ser Cys Ser Pro Lys Glu Glu Ala 565 570 575 Tyr Ile Trp Cys Ala Asp Ser Arg Gln Ile Arg Leu Val Asp Gly Gly 580 585 590 Gly Arg Cys Ser Gly Arg Val Glu Ile Leu Asp Gln Gly Ser Trp Gly 595 600 605 Thr Ile Cys Asp Asp Arg Trp Asp Leu Asp Asp Ala Arg Val Val Cys 610 615 620 Lys Gln Leu Gly Cys Gly Glu Ala Leu Asp Ala Thr Val Ser Ser Phe 625 630 635 640 Phe Gly Thr Gly Ser Gly Pro Ile Trp Leu Asp Glu Val Asn Cys Arg 645 650 655 Gly Glu Glu Ser Gln Val Trp Arg Cys Pro Ser Trp Gly Trp Arg Gln 660 665 670 His Asn Cys Asn His Gln Glu Asp Ala Gly Val Ile Cys Ser Gly Phe 675 680 685 Val Arg Leu Ala Gly Gly Asp Gly Pro Cys Ser Gly Arg Val Glu Val 690 695 700 His Ser Gly Glu Ala Trp Thr Pro Val Ser Asp Gly Asn Phe Thr Leu 705 710 715 720 Pro Thr Ala Gln Val Ile Cys Ala Glu Leu Gly Cys Gly Lys Ala Val 725 730 735 Ser Val Leu Gly His Met Pro Phe Arg Glu Ser Asp Gly Gln Val Trp 740 745 750 Ala Glu Glu Phe Arg Cys Asp Gly Gly Glu Pro Glu Leu Trp Ser Cys 755 760 765 Pro Arg Val Pro Cys Pro Gly Gly Thr Cys Leu His Ser Gly Ala Ala 770 775 780 Gln Val Val Cys Ser Val Tyr Thr Glu Val Gln Leu Met Lys Asn Gly 785 790 795 800 Thr Ser Gln Cys Glu Gly Gln Val Glu Met Lys Ile Ser Gly Arg Trp 805 810 815 Arg Ala Leu Cys Ala Ser His Trp Ser Leu Ala Asn Ala Asn Val Val 820 825 830 Cys Arg Gln Leu Gly Cys Gly Val Ala Ile Ser Thr Pro Arg Gly Pro 835 840 845 His Leu Val Glu Gly Gly Asp Gln Ile Ser Thr Ala Gln Phe His Cys 850 855 860 Ser Gly Ala Glu Ser Phe Leu Trp Ser Cys Pro Val Thr Ala Leu Gly 865 870 875 880 Gly Pro Asp Cys Ser His Gly Asn Thr Ala Ser Val Ile Cys Ser Gly 885 890 895 Asn His Thr Gln Val Leu Pro Gln Cys Asn Asp Phe Leu Ser Gln Pro 900 905 910 Ala Gly Ser Ala Ala Ser Glu Glu Ser Ser Pro Tyr Cys Ser Asp Ser 915 920 925 Arg Gln Leu Arg Leu Val Asp Gly Gly Gly Pro Cys Gly Gly Arg Val 930 935 940 Glu Ile Leu Asp Gln Gly Ser Trp Gly Thr Ile Cys Asp Asp Asp Trp 945 950 955 960 Asp Leu Asp Asp Ala Arg Val Val Cys Arg Gln Leu Gly Cys Gly Glu 965 970 975 Ala Leu Asn Ala Thr Gly Ser Ala His Phe Gly Ala Gly Ser Gly Pro 980 985 990 Ile Trp Leu Asp Asp Leu Asn Cys Thr Gly Lys Glu Ser His Val Trp 995 1000 1005 Arg Cys Pro Ser Arg Gly Trp Gly Arg His Asp Cys Arg His Lys Glu 1010 1015 1020 Asp Ala Gly Val Ile Cys Ser Glu Phe Leu Ala Leu Arg Met Val Ser 1025 1030 1035 1040 Glu Asp Gln Gln Cys Ala Gly Trp Leu Glu Val Phe Tyr Asn Gly Thr 1045 1050 1055 Trp Gly Ser Val Cys Arg Ser Pro Met Glu Asp Ile Thr Val Ser Val 1060 1065 1070 Ile Cys Arg Gln Leu Gly Cys Gly Asp Ser Gly Ser Leu Asn Thr Ser 1075 1080 1085 Val Gly Leu Arg Glu Gly Ser Arg Pro Arg Trp Val Asp Leu Ile Gln 1090 1095 1100 Cys Arg Lys Met Asp Thr Ser Leu Trp Gln Cys Pro Ser Gly Pro Trp 1105 1110 1115 1120 Lys Tyr Ser Ser Cys Ser Pro Lys Glu Glu Ala Tyr Ile Ser Cys Glu 1125 1130 1135 Gly Arg Arg Pro Lys Ser Cys Pro Thr Ala Ala Ala Cys Thr Asp Arg 1140 1145 1150 Glu Lys Leu Arg Leu Arg Gly Gly Asp Ser Glu Cys Ser Gly Arg Val 1155 1160 1165 Glu Val Trp His Asn Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp 1170 1175 1180 Ser Leu Ala Glu Ala Glu Val Val Cys Gln Gln Leu Gly Cys Gly Gln 1185 1190 1195 1200 Ala Leu Glu Ala Val Arg Ser Ala Ala Phe Gly Pro Gly Asn Gly Ser 1205 1210 1215 Ile Trp Leu Asp Glu Val Gln Cys Gly Gly Arg Glu Ser Ser Leu Trp 1220 1225 1230 Asp Cys Val Ala Glu Pro Trp Gly Gln Ser Asp Cys Lys His Glu Glu 1235 1240 1245 Asp Ala Gly Val Arg Cys Ser Gly Val Arg Thr Thr Leu Pro Thr Thr 1250 1255 1260 Thr Ala Gly Thr Arg Thr Thr Ser Asn Ser Leu Pro Gly Ile Phe Ser 1265 1270 1275 1280 Leu Pro Gly Val Leu Cys Leu Ile Leu Gly Ser Leu Leu Phe Leu Val 1285 1290 1295 Leu Val Ile Leu Val Thr Gln Leu Leu Arg Trp Arg Ala Glu Arg Arg 1300 1305 1310 Ala Leu Ser Ser Tyr Glu Asp Ala Leu Ala Glu Ala Val Tyr Glu Glu 1315 1320 1325 Leu Asp Tyr Leu Leu Thr Gln Lys Glu Gly Leu Gly Ser Pro Asp Gln 1330 1335 1340 Met Thr Asp Val Pro Asp Glu Asn Tyr Asp Asp Ala Glu Glu Val Pro 1345 1350 1355 1360 Val Pro Gly Thr Pro Ser Pro Ser Gln Gly Asn Glu Glu Glu Val Pro 1365 1370 1375 Pro Glu Lys Glu Asp Gly Val Arg Ser Ser Gln Thr Gly Ser Phe Leu 1380 1385 1390 Asn Phe Ser Arg Glu Ala Ala Asn Pro Gly Glu Gly Glu Glu Ser Phe 1395 1400 1405 Trp Leu Leu Gln Gly Lys Lys Gly Asp Ala Gly Tyr Asp Asp Val Glu 1410 1415 1420 Leu Ser Ala Leu Gly Thr Ser Pro Val Thr Phe Ser 1425 1430 1435 79 4308 DNA Bos sp. 79 atggctctgg gcagacacct ctccctgcgg ggactctgtg tcctcctcct cggcaccatg 60 gtgggtggtc aagctctgga gctgaggttg aaggatggag tccatcgctg tgaggggaga 120 gtggaagtga agcaccaagg agaatggggc acagtggatg gttacaggtg gacattgaag 180 gatgcatctg tagtgtgcag acagctgggg tgtggagctg ccattggttt tcctggaggg 240 gcttattttg ggccaggact tggccccatt tggcttttgt atacttcatg tgaagggaca 300 gagtcaactg tcagtgactg tgagcattct aatattaaag actatcgtaa tgatggctat 360 aatcatggtc gggatgctgg agtagtctgc tcaggatttg tgcgtctggc tggaggggat 420 ggaccctgct cagggcgagt agaagtgcat tctggagaag cttggatccc agtgtctgat 480 gggaacttca cacttgccac tgcccagatc atctgtgcag agttgggttg tggcaaggct 540 gtgtctgtcc tgggacatga gctcttcaga gagtccagtg cccaggtctg ggctgaagag 600 ttcaggtgtg agggggagga gcctgagctc tgggtctgcc ccagagtgcc ctgtccaggg 660 ggcacgtgtc accacagtgg atctgctcag gttgtttgtt cagcatactc agaagtccgg 720 ctcatgacaa acggctcctc tcagtgtgaa gggcaggtgg agatgaacat ttctggacaa 780 tggagagcgc tctgtgcctc ccactggagt ctggccaatg ccaatgttat ctgtcgtcag 840 ctcggctgtg gagttgccat ctccaccccc ggaggaccac acttggtgga agaaggtgat 900 cagatcctaa cagcccgatt tcactgctct ggggctgagt ccttcctgtg gagttgtcct 960 gtgactgccc tgggtggtcc tgactgttcc catggcaaca cagcctctgt gatctgctca 1020 ggaaaccaga tccaggtgct tccccagtgc aacgactccg tgtctcaacc tacaggctct 1080 gcggcctcag aggacagcgc cccctactgc tcagacagca ggcagctccg cctggtggac 1140 gggggcggtc cctgcgccgg gagagtggag atccttgacc agggctcctg gggcaccatc 1200 tgtgatgacg gctgggacct ggacgatgcc cgcgtggtgt gcaggcagct gggctgtgga 1260 gaagccctca atgccacggg gtctgctcac ttcggggcag gatcagggcc catctggttg 1320 gacaacttga actgcacagg aaaggagtcc cacgtgtgga ggtgcccttc ccggggctgg 1380 gggcagcaca actgcagaca caagcaggac gcgggggtca tctgctcaga gttcctggcc 1440 ctcaggatgg tgagtgagga ccagcagtgt gctgggtggc tggaagtttt ctacaatggg 1500 acctggggca gtgtctgccg taaccccatg gaagacatca ctgtgtccac gatctgcaga 1560 cagcttggct gtggggacag tggaaccctc aactcttctg ttgctcttag agaaggtttt 1620 aggccacagt gggtggatag aatccagtgt cggaaaactg acacctctct ctggcagtgt 1680 ccttctgacc cttggaatta caactcatgc tctccaaagg aggaagccta tatctggtgt 1740 gcagacagca gacagatccg cctggtggat ggaggtggtc gctgctctgg gagagtggag 1800 atccttgacc agggctcctg gggcaccatc tgtgatgacc gctgggacct ggacgatgcc 1860 cgtgtggtgt gcaagcagct gggctgtgga gaagccctgg acgccactgt ctcttccttc 1920 ttcgggacgg gatcagggcc catctggctg gatgaagtga actgcagagg agaggagtcc 1980 caagtatgga ggtgcccttc ctggggatgg cggcaacaca actgcaatca tcaagaagat 2040 gcaggagtca tctgctcagg atttgtgcgt ctggctggag gagatggacc ctgctcaggg 2100 cgagtagaag tgcattctgg agaagcctgg accccagtgt ctgatggaaa cttcacactc 2160 cccactgccc aggtcatctg tgcagagctg ggatgtggca aggctgtgtc tgtcctggga 2220 cacatgccat tcagagagtc cgatggccag gtctgggctg aagagttcag gtgtgatggg 2280 ggggagcctg agctctggtc ctgccccaga gtgccctgtc caggaggcac atgtctccac 2340 agtggagctg ctcaggttgt ctgttcagtg tacacagaag tccagcttat gaaaaacggc 2400 acctctcaat gtgaggggca ggtggagatg aagatctctg gacgatggag agcgctctgt 2460 gcctcccact ggagtctggc caatgccaat gttgtctgtc gtcagctcgg ctgtggagtc 2520 gccatctcca cccccagagg accacacttg gtggaaggag gtgatcagat ctcaacagcc 2580 caatttcact gctcaggggc tgagtccttc ctgtggagtt gtcctgtgac tgccttgggt 2640 gggcctgact gttcccatgg caacacagcc tctgtgatct gctcaggaaa ccacacccag 2700 gtgctgcccc agtgcaacga cttcctgtct caacctgcag gctctgcggc ctcagaggag 2760 agttctccct actgctcaga cagcaggcag ctccgcctgg tggacggggg cggtccctgc 2820 ggcgggagag tggagatcct tgaccagggc tcctggggca ccatctgtga tgatgactgg 2880 gacctggacg atgcccgtgt ggtgtgcagg cagctgggct gtggagaagc cctcaatgcc 2940 acggggtctg ctcacttcgg ggcaggatca gggcccatct ggctggacga cctgaactgc 3000 acaggaaagg agtcccacgt gtggaggtgc ccttcccggg gctgggggcg gcacgactgc 3060 agacacaagg aggacgccgg ggtcatctgc tcagagttcc tggccctcag gatggtgagc 3120 gaggaccagc agtgtgctgg gtggctggag gttttctaca acgggacctg gggcagtgtc 3180 tgccgcagcc ccatggaaga tatcactgtg tccgtgatct gcagacagct tggatgtggg 3240 gacagtggaa gtctcaacac ctctgttggt ctcagggaag gttctagacc ccggtgggta 3300 gatttaattc agtgtcggaa aatggatacc tctctctggc agtgtccttc tggcccatgg 3360 aaatacagtt catgctctcc aaaggaggaa gcctacatct catgtgaagg aagaagaccc 3420 aagagctgtc caactgctgc cgcctgcaca gacagagaga agctccgcct caggggagga 3480 gacagcgagt gctcagggcg ggtggaggtg tggcacaacg gctcctgggg caccgtgtgc 3540 gatgactcct ggagcctggc agaggctgag gtggtgtgtc agcagctggg ctgtggccag 3600 gccctggaag ccgtgcggtc tgcagcattt ggccctggaa atgggagcat ctggctggac 3660 gaggtgcagt gcgggggccg ggagtcctcc ctgtgggact gtgttgcgga gccctggggg 3720 cagagcgact gcaagcacga ggaggatgct ggtgtgaggt gctctggtgt aaggacaaca 3780 ttgcccacga ccacagcagg gaccagaaca acctcaaatt ctctccctgg catcttctcc 3840 ctgcctgggg ttctctgcct tatcctgggg tcgcttctct tcctggtcct cgtcatcctg 3900 gtgactcagc tactcagatg gagagcagag cgcagagcct tatccagcta tgaagatgct 3960 cttgctgaag ctgtgtatga ggagctcgat taccttctga cacagaagga aggtctgggc 4020 agcccagatc agatgactga tgtccctgat gaaaattatg atgatgctga agaagtacca 4080 gtgcctggaa ctccttctcc ctctcagggg aatgaggagg aagtgccccc agagaaggag 4140 gacggggtga ggtcctctca gacaggctct ttcctgaact tctccagaga ggcagctaat 4200 cctggggaag gagaagagag cttctggctg ctccagggga agaaagggga tgctgggtat 4260 gatgatgttg aactcagtgc cctgggaaca tccccagtga ctttctcg 4308 

What is claimed is:
 1. An isolated polypeptide selected from the group consisting of: a) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO:45 or SEQ ID NO:46; b) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 90% identical to the nucleotide sequence of the DNA insert of clone EpT294, which was deposited with ATCC as Accession Number 207220; c) a polypeptide which is at least 90% identical to the amino acid sequence of SEQ ID NO:47; d) a polypeptide which is at least 90% identical to residues 15-423 of the amino acid sequence of SEQ ID NO:47; and e) a polypeptide which is at least 90% identical to the amino acid sequence of SEQ ID NO: 49; wherein the polypeptide exhibits a lipase activity.
 2. The isolated polypeptide of claim 1, wherein the polypeptide is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the nucleotide sequence of SEQ ID NO:45 or SEQ ID NO:46.
 3. The isolated polypeptide of claim 1, wherein the polypeptide is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the nucleotide sequence of the DNA insert of clone EpT294, which was deposited with ATCC as Accession Number
 207220. 4. The isolated polypeptide of claim 1, wherein the polypeptide is at least 95% identical to the amino acid sequence of SEQ ID NO:47.
 5. The isolated polypeptide of claim 1, wherein the polypeptide is at least 95% identical to residues 15-423 of the amino acid sequence of SEQ ID NO:47.
 6. The isolated polypeptide of claim 1, wherein the polypeptide is at least 95% identical to the amino acid sequence of SEQ ID NO:49.
 7. The isolated polypeptide of claim 1, wherein the amino acid sequence of the polypeptide further comprises heterologous amino acid residues.
 8. The isolated polypeptide of claim 1, admixed with a pharmaceutically acceptable carrier.
 9. An isolated polypeptide which is encoded by a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:45 or SEQ ID NO:46.
 10. The isolated polypeptide of claim 9, wherein the amino acid sequence of the polypeptide further comprises heterologous amino acid residues.
 11. The isolated polypeptide of claim 9, admixed with a pharmaceutically acceptable carrier.
 12. An isolated polypeptide which is encoded by the nucleotide sequence of the DNA insert of clone EpT294, which was deposited with ATCC as Accession Number
 207220. 13. The isolated polypeptide of claim 12, wherein the amino acid sequence of the polypeptide further comprises heterologous amino acid residues.
 14. The isolated polypeptide of claim 12, admixed with a pharmaceutically acceptable carrier.
 15. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO:47.
 16. The isolated polypeptide of claim 15, wherein the amino acid sequence of the polypeptide further comprises heterologous amino acid residues.
 17. The isolated polypeptide of claim 15, admixed with a pharmaceutically acceptable carrier.
 18. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO:49.
 19. The isolated polypeptide of claim 18, wherein the amino acid sequence of the polypeptide further comprises heterologous amino acid residues.
 20. The isolated polypeptide of claim 18, admixed with a pharmaceutically acceptable carrier. 